2024/02/15 更新

写真b

タチカワ ヒロユキ
舘川 宏之
TACHIKAWA Hiroyuki
*大学が定期的に情報更新している項目(その他は、researchmapの登録情報を転載)
所属*
スポーツウエルネス学部 スポーツウエルネス学科
スポーツウエルネス学研究科 スポーツウエルネス学専攻 博士課程前期課程
スポーツウエルネス学研究科 スポーツウエルネス学専攻 博士課程後期課程
職名*
教授
学位
博士(農学) ( 東京大学 )
研究テーマ*
  • スポーツ、健康、老化、疾患は、広い意味でマクロなレベルのヒトの生命現象であり、その理解には遺伝子や細胞などのミクロなレベルからの視点がこれから一層重要となる。私達は、培養細胞や酵母を細胞のモデルとして用いて、細胞内で起きるさまざまな現象のメカニズム解明を目指している。特に、細胞内のオルガネラに着目し、膜動態や形成メカニズム、生化学的機能の解明を通して、ヒトの生命現象の理解へとつながる研究を行っている。

  • 研究キーワード
  • 脂質輸送

  • 小胞輸送

  • 胞子形成

  • 出芽酵母

  • 膜形成

  • 生体膜

  • 学内職務経歴*
    • 2023年4月 - 現在 
      スポーツウエルネス学部   スポーツウエルネス学科   教授
    • 2023年4月 - 現在 
      スポーツウエルネス学研究科   スポーツウエルネス学専攻 博士課程前期課程   教授
    • 2023年4月 - 現在 
      スポーツウエルネス学研究科   スポーツウエルネス学専攻 博士課程後期課程   教授
     

    研究分野

    • ライフサイエンス / 応用分子細胞生物学

    • ライフサイエンス / 応用微生物学

    経歴

    • 2023年4月 - 現在 
      立教大学   スポーツウエルネス学部   教授

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    • 2009年10月 - 2023年3月 
      東京大学   農学生命科学研究科   准教授

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    • 2004年10月 - 2009年9月 
      東京大学   農学生命科学研究科   講師

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    • 1993年4月 - 2004年9月 
      東京農工大学   応用生物科学科   助手

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    学歴

    • 1990年4月 - 1993年3月 
      東京大学大学院   農学系研究科   農芸化学専攻博士課程

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    • 1988年4月 - 1990年3月 
      東京大学大学院   農学系研究科   農芸化学専攻修士課程

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    • - 1988年3月 
      東京大学   農学部   農芸化学科

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    委員歴

    • 2019年12月 - 2021年9月 
      酵母細胞研究会   第23回酵母合同シンポジウム実行委員会委員

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    • 2019年9月 - 2020年5月 
      日本農芸化学会   本部選挙管理委員会委員

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    • 2017年12月 - 2020年2月 
      日本農芸化学会   2019年度東京大会実行委員会委員

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      団体区分:学協会

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    • 2017年9月 - 2018年5月 
      日本農芸化学会   本部選挙管理委員会委員

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      団体区分:学協会

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    • 2016年12月 - 2017年10月 
      酵母遺伝学フォーラム   2017年度研究報告会実行委員

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      団体区分:学協会

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    • 2013年3月 - 2015年2月 
      日本農芸化学会   2014年度東京大会実行委員会委員

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      団体区分:学協会

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    • 2013年12月 - 2014年9月 
      酵母細胞研究会   第21回酵母合同シンポジウム実行委員会委員

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      団体区分:その他

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    ▼全件表示

    論文

    • Application of yeast spores as β-glucan particles

      Guoyu Liu, Yan Yang, Hiroyuki Tachikawa, Xiao-Dong Gao, Hideki Nakanishi

      Particuology71   34 - 40   2022年12月

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      掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

      DOI: 10.1016/j.partic.2022.01.013

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    • Receptor for advanced glycation end-products (RAGE) mediates phagocytosis in nonprofessional phagocytes. 国際誌

      Yan Yang, Guoyu Liu, Feng Li, Lucas B Carey, Changjin Sun, Kaiping Ling, Hiroyuki Tachikawa, Morihisa Fujita, Xiao-Dong Gao, Hideki Nakanishi

      Communications biology5 ( 1 ) 824 - 824   2022年8月16日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      In mammals, both professional phagocytes and nonprofessional phagocytes (NPPs) can perform phagocytosis. However, limited targets are phagocytosed by NPPs, and thus, the mechanism remains unclear. We find that spores of the yeast Saccharomyces cerevisiae are internalized efficiently by NPPs. Analyses of this phenomenon reveals that RNA fragments derived from cytosolic RNA species are attached to the spore wall, and these fragments serve as ligands to induce spore internalization. Furthermore, we show that a multiligand receptor, RAGE (receptor for advanced glycation end-products), mediates phagocytosis in NPPs. RAGE-mediated phagocytosis is not uniquely induced by spores but is an intrinsic mechanism by which NPPs internalize macromolecules containing RAGE ligands. In fact, artificial particles labeled with polynucleotides, HMGB1, or histone (but not bovine serum albumin) are internalized in NPPs. Our findings provide insight into the molecular basis of phagocytosis by NPPs, a process by which a variety of macromolecules are targeted for internalization.

      DOI: 10.1038/s42003-022-03791-1

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    • Suppression of Vps13 adaptor protein mutants reveals a central role for PI4P in regulating prospore membrane extension. 国際誌

      Tsuyoshi S Nakamura, Yasuyuki Suda, Kenji Muneshige, Yuji Fujieda, Yuuya Okumura, Ichiro Inoue, Takayuki Tanaka, Tetsuo Takahashi, Hideki Nakanishi, Xiao-Dong Gao, Yasushi Okada, Aaron M Neiman, Hiroyuki Tachikawa

      PLoS genetics17 ( 8 ) e1009727   2021年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      Vps13 family proteins are proposed to function in bulk lipid transfer between membranes, but little is known about their regulation. During sporulation of Saccharomyces cerevisiae, Vps13 localizes to the prospore membrane (PSM) via the Spo71-Spo73 adaptor complex. We previously reported that loss of any of these proteins causes PSM extension and subsequent sporulation defects, yet their precise function remains unclear. Here, we performed a genetic screen and identified genes coding for a fragment of phosphatidylinositol (PI) 4-kinase catalytic subunit and PI 4-kinase noncatalytic subunit as multicopy suppressors of spo73Δ. Further genetic and cytological analyses revealed that lowering PI4P levels in the PSM rescues the spo73Δ defects. Furthermore, overexpression of VPS13 and lowering PI4P levels synergistically rescued the defect of a spo71Δ spo73Δ double mutant, suggesting that PI4P might regulate Vps13 function. In addition, we show that an N-terminal fragment of Vps13 has affinity for the endoplasmic reticulum (ER), and ER-plasma membrane (PM) tethers localize along the PSM in a manner dependent on Vps13 and the adaptor complex. These observations suggest that Vps13 and the adaptor complex recruit ER-PM tethers to ER-PSM contact sites. Our analysis revealed that involvement of a phosphoinositide, PI4P, in regulation of Vps13, and also suggest that distinct contact site proteins function cooperatively to promote de novo membrane formation.

      DOI: 10.1371/journal.pgen.1009727

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    • Identification of novel O-GlcNAc transferase substrates using yeast cells expressing OGT.

      Feng Li, Ganglong Yang, Hiroyuki Tachikawa, Kankai Shao, Yan Yang, Xiao-Dong Gao, Hideki Nakanishi

      The Journal of general and applied microbiology67 ( 1 ) 33 - 41   2021年4月16日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      O-GlcNAc modification mediated by O-GlcNAc transferase (OGT) is a reversible protein modification in which O-GlcNAc moieties are attached to target proteins in the cytosol, nucleus, and mitochondria. O-GlcNAc moieties attached to proteins can be removed by O-GlcNAcase (OGA). The addition of an O-GlcNAc moiety can influence several aspects of protein function, and aberrant O-GlcNAc modification is linked to a number of diseases. While OGT and OGA are conserved across eukaryotic cells, yeasts lack these enzymes. Previously, we reported that protein O-GlcNAc modification occurred in the budding yeast Saccharomyces cerevisiae when OGT was ectopically expressed. Because yeast cells lack OGA, O-GlcNAc moieties are stably attached to target proteins. Thus, the yeast system may be useful for finding novel OST substrates. By proteomic analysis, we identified 468 O-GlcNAcylated proteins in yeast cells expressing human OGT. Among these proteins, 13 have human orthologues that show more than 30% identity to their corresponding yeast orthologue, and possible glycosylation residues are conserved in these human orthologues. In addition, the orthologues have not been reported as substrates of OGT. We verified that some of these human orthologues are O-GlcNAcylated in cultured human cells. These proteins include an ubiquitin-conjugating enzyme, UBE2D1, and an eRF3-similar protein, HBS1L. Thus, the yeast system would be useful to find previously unknown O-GlcNAcylated proteins and regulatory mechanisms.

      DOI: 10.2323/jgam.2020.04.002

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    • LYAR potentiates rRNA synthesis by recruting BRD2/4 and MYST-type acetyltransferase KAT7 to rDNA 査読有り 国際誌

      Nucleic Acids Research47 ( 19 ) 10357 - 10372   2019年11月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1093/nar/gkz747

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    • Osw2 is required for proper assembly of glucan and/or mannan layers of the yeast spore wall 査読有り

      Hua-Ping Pan, Ning Wang, Hiroyuki Tachikawa, Xiao-Dong Gao, Hideki Nakanishi

      Journal of Biochemistry163 ( 4 ) 293 - 304   2018年4月1日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Oxford University Press  

      OSW2 is a meiotically-induced gene required for spore wall formation. osw2Δspores are sensitive to ether treatment. Except for this phenotype, the mutants do not show obvious sporulation defects
      thus, its function remains elusive. We found that deletion of both OSW2 and CHS3 results in a synthetic sporulation defect. The spore wall is composed of four layers, and chs3Δspores lack the outer two (chitosan and dityrosine) layers. Thus, Osw2 is involved in the assembly of the inner (glucan and mannan) layers. In agreement with this notion, a glycosylphosphatidylinositol-anchored protein reporter mislocalizes in osw2Δspores. The osw2Δmutation also exhibited a severe synthetic sporulation defect when combined with the deletion of a ß-1,6-glucan synthesis-related gene, BIG1. Osw2 is localized to the prospore membrane during sporulation. However, it disappears in mature spores, indicating that it is not a structural component of the spore wall. Given that Osw2 contains a probable 2-dehydropantoate 2-reductase domain, it may mediate an enzymatic reaction. Osw2 shows a weak similarity to other 2-dehydropantoate 2-reductase domain-containing proteins, Svl3 and Pam1. A pam1"svl3Δmutant exhibits vegetative cell and spore wall defects. Thus, the 2-dehydropantoate 2-reductase domain-containing proteins may have a similar function in glucan and/or mannan layer assembly.

      DOI: 10.1093/jb/mvx082

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    • Activation of Rab GTPase Sec4 by its GEF Sec2 is required for prospore membrane formation during sporulation in yeast Saccharomyces cerevisiae. 査読有り

      Suda Y, Tachikawa H, Inoue I, Kurita T, Saito C, Kurokawa K, Nakano A, Irie K

      FEMS Yeast Research18 ( 1 )   2018年2月

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      掲載種別:研究論文(学術雑誌)  

      DOI: 10.1093/femsyr/fox095

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    • Dynamic localization of a yeast development-specific PP1 complex during prospore membrane formation is dependent on multiple localization signals and complex formation 査読有り

      Tsuyoshi S. Nakamura, Yumi Numajiri, Yuuya Okumura, Junji Hidaka, Takayuki Tanaka, Ichiro Inoue, Yasuyuki Suda, Tetsuo Takahashi, Hideki Nakanishi, Xiao-Dong Gao, Aaron M. Neiman, Hiroyuki Tachikawa

      MOLECULAR BIOLOGY OF THE CELL28 ( 26 ) 3881 - 3895   2017年12月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC CELL BIOLOGY  

      During the developmental process of sporulation in Saccharomyces cerevisiae, membrane structures called prospore membranes are formed de novo, expand, extend, acquire a round shape, and finally become plasma membranes of the spores. GIP1 encodes a regulatory/targeting subunit of protein phosphatase type 1 that is required for sporulation. Gip1 recruits the catalytic subunit Glc7 to septin structures that form along the prospore membrane; however, the molecular basis of its localization and function is not fully understood. Here we show that Gip1 changes its localization dynamically and is required for prospore membrane extension. Gip1 first associates with the spindle pole body as the prospore membrane forms, moves onto the prospore membrane and then to the septins as the membrane extends, distributes around the prospore membrane after closure, and finally translocates into the nucleus in the maturing spore. Deletion and mutation analyses reveal distinct sequences in Gip1 that are required for different localizations and for association with Glc7. Binding to Glc7 is also required for proper localization. Strikingly, localization to the prospore membrane, but not association with septins, is important for Gip1 function. Further, our genetic analysis suggests that a Gip1-Glc7 phosphatase complex regulates prospore membrane extension in parallel to the previously reported Vps13, Spo71, Spo73 pathway.

      DOI: 10.1091/mbc.E17-08-0521

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    • β-1,6-glucan synthesis-associated genes are required for proper spore wall formation in Saccharomyces cerevisiae. 査読有り

      Pan HP, Wang N, Tachikawa H, Nakanishi H, Gao XD

      Yeast34 ( 11 ) 431 - 446   2017年11月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1002/yea.3244

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    • Identification and characterization of transcriptional control region of the human beta 1,4-mannosyltransferase gene 査読有り

      Tetsuo Takahashi, Takashi Nedachi, Takuya Etoh, Hiroyuki Tachikawa, Xiao-Dong Gao

      CYTOTECHNOLOGY69 ( 3 ) 417 - 434   2017年6月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

      All asparagine-linked glycans (N-glycans) on the eukaryotic glycoproteins are primarily derived from dolichol-linked oligosaccharides (DLO), synthesized on the rough endoplasmic reticulum membrane. We have previously reported cloning and identification of the human gene, HMT-1, which encodes chitobiosyldiphosphodolichol beta-mannosyltransferase (beta 1,4-MT) involved in the early assembly of DLO. Considering that N-glycosylation is one of the most ubiquitous post-translational modifications for many eukaryotic proteins, the HMT-1 could be postulated as one of the housekeeping genes, but its transcriptional regulation remains to be investigated. Here we screened a 1 kb region upstream from HMT-1 open reading frame (ORF) for transcriptionally regulatory sequences by using chloramphenicol acetyl transferase (CAT) assay, and found that the region from -33 to -1 positions might act in HMT-1 transcription at basal level and that the region from -200 to -42 should regulate its transcription either positively or negatively. In addition, results with CAT assays suggested the possibility that two GATA-1 motifs and an Sp1 motif within a 200 bp region upstream from HMT-1 ORF might significantly upregulate HMT-1 transcription. On the contrary, the observations obtained from site-directed mutational analyses revealed that an NF-1/AP-2 overlapping motif located at -148 to -134 positions should serve as a strong silencer. The control of the HMT-1 transcription by these motifs resided within the 200 bp region could partially explain the variation of expression level among various human tissues, suggesting availability and importance of this region for regulatory role in HMT-1 expression.

      DOI: 10.1007/s10616-015-9929-y

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    • The Dysferlin Domain-Only Protein, Spo73, Is Required for Prospore Membrane Extension in Saccharomyces cerevisiae 査読有り

      Yuuya Okumura, Tsuyoshi S. Nakamura, Takayuki Tanaka, Ichiro Inoue, Yasuyuki Suda, Tetsuo Takahashi, Hideki Nakanishi, Shugo Nakamura, Xiao-Dong Gao, Hiroyuki Tachikawa

      MSPHERE1 ( 1 )   2016年1月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

      Sporulation of Saccharomyces cerevisiae is a developmental process in which an ascus containing four haploid spores forms from a diploid cell. During this process, newly formed membrane structures called prospore membranes extend along the nuclear envelope and engulf and package daughter nuclei along with cytosol and organelles to form precursors of spores. Proteins involved in prospore membrane extension, Vps13 and Spo71, have recently been reported; however, the overall mechanism of membrane extension remains unclear. Here, we identified Spo73 as an additional factor involved in prospore membrane extension. Analysis of a spo73 Delta mutant revealed that it shows defects similar to those of a spo71 Delta mutant during prospore membrane formation. Spo73 localizes to the prospore membrane, and this localization is independent of Spo71 and Vps13. In contrast, a Spo73 protein carrying mutations in a surface basic patch mislocalizes to the cytoplasm and overexpression of Spo71 can partially rescue localization to the prospore membrane. Similar to spo71 Delta mutants, spo73 Delta mutants display genetic interactions with the mutations in the SMA2 and SPO1 genes involved in prospore membrane bending. Further, our bioinformatic analysis revealed that Spo73 is a dysferlin domain-only protein. Thus, these results suggest that a dysferlin domain-only protein, Spo73, functions with a dual pleckstrin homology domain protein, Spo71, in prospore membrane extension. Analysis of Spo73 will provide insights into the conserved function of dysferlin domains, which is related to dysferlinopathy.
      IMPORTANCE Prospore membrane formation consists of de novo double-membrane formation, which occurs during the developmental process of sporulation in Saccharomyces cerevisiae. Membranes are formed into their proper size and shape, and thus, prospore membrane formation has been studied as a general model of membrane formation. We identified SPO73, previously shown to be required for spore wall formation, as an additional gene involved in prospore membrane extension. Genetic and cell biological analyses suggested that Spo73 functions on the prospore membrane with other factors in prospore membrane extension, counteracting the bending force of the prospore membrane. Spo73 is the first dysferlin domain-only protein ever analyzed. The dysferlin domain is conserved from yeast to mammals and is found in dysferlin proteins, which are involved in dysferlinopathy, although the precise function of the domain is unknown. Continued analysis of Spo73 will contribute to our understanding of the function of dysferlin domains and dysferlinopathy.

      DOI: 10.1128/mSphere.00038-15

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    • Yeast cell-based analysis of human lactate dehydrogenase isoforms 査読有り

      Lulu Ahmed Mohamed, Hiroyuki Tachikawa, Xiao-Dong Gao, Hideki Nakanishi

      JOURNAL OF BIOCHEMISTRY158 ( 6 ) 467 - 476   2015年12月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

      Human lactate dehydrogenase (LDH) has attracted attention as a potential target for cancer therapy and contraception. In this study, we reconstituted human lactic acid fermentation in Saccharomyces cerevisiae, with the goal of constructing a yeast cell-based LDH assay system. pdc null mutant yeast (mutated in the endogenous pyruvate decarboxylase genes) are unable to perform alcoholic fermentation; when grown in the presence of an electron transport chain inhibitor, pdc null strains exhibit a growth defect. We found that introduction of the human gene encoding LDHA complemented the pdc growth defect; this complementation depended on LDHA catalytic activity. Similarly, introduction of the human LDHC complemented the pdc growth defect, even though LDHC did not generate lactate at the levels seen with LDHA. In contrast, the human LDHB did not complement the yeast pdc null mutant, although LDHB did generate lactate in yeast cells. Expression of LDHB as a red fluorescent protein (RFP) fusion yielded blebs in yeast, whereas LDHA-RFP and LDHC-RFP fusion proteins exhibited cytosolic distribution. Thus, LDHB exhibits several unique features when expressed in yeast cells. Because yeast cells are amenable to genetic analysis and cell-based high-throughput screening, our pdc/LDH strains are expected to be of use for versatile analyses of human LDH.

      DOI: 10.1093/jb/mvv061

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    • Use of Yeast Spores for Microencapsulation of Enzymes 査読有り

      Libing Shi, Zijie Li, Hiroyuki Tachikawa, Xiao-Dong Gao, Hideki Nakanishi

      APPLIED AND ENVIRONMENTAL MICROBIOLOGY80 ( 15 ) 4502 - 4510   2014年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

      Here, we report a novel method to produce microencapsulated enzymes using Saccharomyces cerevisiae spores. In sporulating cells, soluble secreted proteins are transported to the spore wall. Previous work has shown that the spore wall is capable of retaining soluble proteins because its outer layers work as a diffusion barrier. Accordingly, a red fluorescent protein (RFP) fusion of the alpha-galactosidase, Mel1, expressed in spores was observed in the spore wall even after spores were subjected to a high-salt wash in the presence of detergent. In vegetative cells, however, the cell wall cannot retain the RFP fusion. Although the spore wall prevents diffusion of proteins, it is likely that smaller molecules, such as sugars, pass through it. In fact, spores can contain much higher alpha-galactosidase activity to digest melibiose than vegetative cells. When present in the spore wall, the enzyme acquires resistance to environmental stresses including enzymatic digestion and high temperatures. The outer layers of the spore wall are required to retain enzymes but also decrease accessibility of the substrates. However, mutants with mild spore wall defects can retain and stabilize the enzyme while still permitting access to the substrate. In addition to Mel1, we also show that spores can retain the invertase. Interestingly the encapsulated invertase has significantly lower activity toward raffinose than toward sucrose. This suggests that substrate selectivity could be altered by the encapsulation.

      DOI: 10.1128/AEM.00153-14

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    • Applied Usage of Yeast Spores as Chitosan Beads 査読有り

      Haini Zhang, Hiroyuki Tachikawa, Xiao-Dong Gao, Hideki Nakanishi

      APPLIED AND ENVIRONMENTAL MICROBIOLOGY80 ( 16 ) 5098 - 5105   2014年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

      In this study, we present a nonhazardous biological method of producing chitosan beads using the budding yeast Saccharomyces cerevisiae. Yeast cells cultured under conditions of nutritional starvation cease vegetative growth and instead form spores. The spore wall has a multilaminar structure with the chitosan layer as the second outermost layer. Thus, removal of the outermost dityrosine layer by disruption of the DIT1 gene, which is required for dityrosine synthesis, leads to exposure of the chitosan layer at the spore surface. In this way, spores can be made to resemble chitosan beads. Chitosan has adsorptive features and can be used to remove heavy metals and negatively charged molecules from solution. Consistent with this practical application, we find that spores are capable of adsorbing heavy metals such as Cu2+, Cr3+, and Cd2+, and removal of the dityrosine layer further improves the adsorption. Removal of the chitosan layer decreases the adsorption, indicating that chitosan works as an adsorbent in the spores. Besides heavy metals, spores can also adsorb a negatively charged cholesterol derivative, taurocholic acid. Furthermore, chitosan is amenable to chemical modifications, and, consistent with this property, dit1 Delta spores can serve as a carrier for immobilization of enzymes. Given that yeast spores are a natural product, our results demonstrate that they, and especially dit1 Delta mutants, can be used as chitosan beads and used for multiple purposes.

      DOI: 10.1128/AEM.00677-14

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    • Natural IgG antibody with anti-β-galactosyl specificity suppressed hepatoma cell invasion in culture. 査読有り

      Miura Y, Fujita H, Sakai F, Tachikawa H, Yagasaki K, Fujimoto D

      Cytotechnology65 ( 6 ) 909 - 913   2013年12月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1007/s10616-012-9523-5

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    • SPO71 encodes a developmental stage-specific partner for Vps13 in Saccharomyces cerevisiae 査読有り

      Jae-Sook Park, Yuuya Okumura, Hiroyuki Tachikawa, Aaron M. Neiman

      Eukaryotic Cell12 ( 11 ) 1530 - 1537   2013年11月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      The creation of haploid gametes in yeast, termed spores, requires the de novo formation of membranes within the cytoplasm. These membranes, called prospore membranes, enclose the daughter nuclei generated by meiosis. Proper growth and closure of prospore membranes require the highly conserved Vps13 protein. Mutation of SPO71, a meiosis-specific gene first identified as defective in spore formation, was found to display defects in membrane morphogenesis very similar to those seen in vps13Δ cells. Specifically, prospore membranes are smaller than in the wild type, they fail to close, and membrane vesicles are present within the prospore membrane lumen. As in vps13Δ cells, the levels of phophatidylinositol-4-phosphate are reduced in the prospore membranes of spo71 cells. SPO71Δ is required for the translocation of Vps13 from the endosome to the prospore membrane, and ectopic expression of SPO71 in vegetative cells results in mislocalization of Vps13. Finally, the two proteins can be coprecipitated from sporulating cells. We propose that Spo71 is a sporulation-specific partner for Vps13 and that they act in concert to regulate prospore membrane morphogenesis. © 2013, American Society for Microbiology. All Rights Reserved.

      DOI: 10.1128/EC.00239-13

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    • Alg14 organizes the formation of a multiglycosyltransferase complex involved in initiation of lipid-linked oligosaccharide biosynthesis 査読有り

      Jishun Lu, Tetsuo Takahashi, Atsuko Ohoka, Kei-ichi Nakajima, Ryo Hashimoto, Nobuaki Miura, Hiroyuki Tachikawa, Xiao-Dong Gao

      GLYCOBIOLOGY22 ( 4 ) 504 - 516   2012年4月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS INC  

      Protein N-glycosylation begins with the assembly of a lipid-linked oligosaccharide (LLO) on the endoplasmic reticulum (ER) membrane. The first two steps of LLO biosynthesis are catalyzed by a functional multienzyme complex comprised of the Alg7 GlcNAc phosphotransferase and the heterodimeric Alg13/Alg14 UDP-GlcNAc transferase on the cytosolic face of the ER. In the Alg13/14 glycosyltransferase, Alg14 recruits cytosolic Alg13 to the ER membrane through interaction between their C-termini. Bioinformatic analysis revealed that eukaryotic Alg14 contains an evolved N-terminal region that is missing in bacterial orthologs. Here, we show that this N-terminal region of Saccharomyces cerevisiae Alg14 localize its green fluorescent protein fusion to the ER membrane. Deletion of this region causes defective growth at 38.5 degrees C that can be partially complemented by overexpression of Alg7. Coimmunoprecipitation demonstrated that the N-terminal region of Alg14 is required for direct interaction with Alg7. Our data also show that Alg14 lacking the N-terminal region remains on the ER membrane through a nonperipheral association, suggesting the existence of another membrane-binding site. Mutational studies guided by the 3D structure of Alg14 identified a conserved alpha-helix involved in the second membrane association site that contributes to an integral interaction and protein stability. We propose a model in which the N- and C-termini of Alg14 coordinate recruitment of catalytic Alg7 and Alg13 to the ER membrane for initiating LLO biosynthesis.

      DOI: 10.1093/glycob/cwr162

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    • Splicing Factor 2-Associated Protein p32 Participates in Ribosome Biogenesis by Regulating the Binding of Nop52 and Fibrillarin to Preribosome Particles 査読有り

      Harunori Yoshikawa, Wataru Komatsu, Toshiya Hayano, Yutaka Miura, Keiichi Homma, Keiichi Izumikawa, Hideaki Ishikawa, Naoki Miyazawa, Hiroyuki Tachikawa, Yoshio Yamauchi, Toshiaki Isobe, Nobuhiro Takahashi

      MOLECULAR & CELLULAR PROTEOMICS10 ( 8 ) M110.006148   2011年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

      Ribosome biogenesis starts with transcription of the large ribosomal RNA precursor (47S pre-rRNA), which soon combines with numerous factors to form the 90S preribosome in the nucleolus. Although the subsequent separation of the pre-90S particle into pre-40S and pre-60S particles is critical for the production process of mature small and large ribosomal subunits, its molecular mechanisms remain undetermined. Here, we present evidence that p32, fibrillarin (FBL), and Nop52 play key roles in this separation step. Mass-based analyses combined with immunoblotting showed that p32 associated with 155 proteins including 31 rRNA-processing factors (of which nine were components of small subunit processome, and six were those of RIX1 complex), 13 chromatin remodeling components, and six general transcription factors required for RNA polymerase III-mediated transcription. Of these, a late rRNA-processing factor Nop52 interacted directly with p32. Immunocytochemical analyses demonstrated that p32 colocalized with an early rRNA-processing factor FBL or Nop52 in the nucleolus and Cajal bodies, but was excluded from the nucleolus after actinomycin D treatment. p32 was present in the pre-ribosomal fractions prepared by cell fractionation or separated by ultracentrifugation of the nuclear extract. p32 also associated with pre-rRNAs including 47S/45S and 32S pre-rRNAs. Furthermore, knockdown of p32 with a small interfering RNA slowed the early processing from 47S/45S pre-rRNAs to 18S rRNA and 32S pre-rRNA. Finally, Nop52 was found to compete with FBL for binding to p32 probably in the nucleolus. Given the fact that FBL and Nop52 are associated with pre-ribosome particles distinctly different from each other, we suggest that p32 is a new rRNA maturation factor involved in the remodeling from pre-90S particles to pre-40S and pre-60S particles that requires the exchange of FBL for Nop52. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.006148, 1-23, 2011.

      DOI: 10.1074/mcp.M110.006148

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    • Role of septins in the orientation of forespore membrane extension during sporulation in fission yeast 査読有り

      Onishi, M., Koga, T., Hirata, A., Nakamura, T., Asakawa, H., Shimoda, C., Bähler, J., Wu, J.-Q., Takegawa, K., Tachikawa, H., Pringle, J.R., Fukui, Y.

      Molecular and Cellular Biology30 ( 8 ) 2057 - 2074   2010年

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      掲載種別:研究論文(学術雑誌)  

      DOI: 10.1128/MCB.01529-09

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    • Protein Phosphatase Type 1-Interacting Protein Ysw1 Is Involved in Proper Septin Organization and Prospore Membrane Formation during Sporulation 査読有り

      Makoto Ishihara, Yasuyuki Suda, Ichiro Inoue, Takayuki Tanaka, Tetsuo Takahashi, Xiao-Dong Gao, Yasuhisa Fukui, Sayoko Ihara, Aaron M. Neiman, Hiroyuki Tachikawa

      EUKARYOTIC CELL8 ( 7 ) 1027 - 1037   2009年7月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

      Sporulation of Saccharomyces cerevisiae is a developmental process in which four haploid spores are generated inside a diploid cell. Gip1, a sporulation-specific targeting subunit of protein phosphatase type 1, together with its catalytic subunit, Glc7, colocalizes with septins along the extending prospore membrane and is required for septin organization and spore wall formation. However, the mechanism by which Gip1-Glc7 phosphatase promotes these events is unclear. We show here that Ysw1, a sporulation-specific coiled-coil protein, has a functional relationship to Gip1-Glc7 phosphatase. Overexpression of YSW1 partially suppresses the sporulation defect of a temperature-sensitive allele of gip1. Ysw1 interacts with Gip1 in a two-hybrid assay, and this interaction is required for suppression. Ysw1 tagged with green fluorescent protein colocalizes with septins and Gip1 along the extending prospore membrane during spore formation. Sporulation is partially defective in ysw1 Delta mutant, and cytological analysis revealed that septin structures are perturbed and prospore membrane extension is aberrant in ysw1 Delta cells. These results suggest that Ysw1 functions with the Gip1-Glc7 phosphatase to promote proper septin organization and prospore membrane formation.

      DOI: 10.1128/EC.00095-09

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    • The Anaphase Promoting Complex Targeting Subunit Ama1 Links Meiotic Exit to Cytokinesis during Sporulation in Saccharomyces cerevisiae 査読有り

      Aviva E. Diamond, Jae-Sook Park, Ichiro Inoue, Hiroyuki Tachikawa, Aaron M. Neiman

      MOLECULAR BIOLOGY OF THE CELL20 ( 1 ) 134 - 145   2009年1月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC CELL BIOLOGY  

      Ascospore formation in yeast is accomplished through a cell division in which daughter nuclei are engulfed by newly formed plasma membranes, termed prospore membranes. Closure of the prospore membrane must be coordinated with the end of meiosis II to ensure proper cell division. AMA1 encodes a meiosis-specific activator of the anaphase promoting complex (APC). The activity of APC(Ama1) is inhibited before meiosis II, but the substrates specifically targeted for degradation by Ama1 at the end of meiosis are unknown. We show here that ama1 Delta mutants are defective in prospore membrane closure. Ssp1, a protein found at the leading edge of the prospore membrane, is stabilized in ama1 Delta mutants. Inactivation of a conditional form of Ssp1 can partially rescue the sporulation defect of the ama1 Delta mutant, indicating that an essential function of Ama1 is to lead to the removal of Ssp1. Depletion of Cdc15 causes a defect in meiotic exit. We find that prospore membrane closure is also defective in Cdc15 and that this defect can be overcome by expression of a form of Ama1 in which multiple consensus cyclin-dependent kinase phosphorylation sites have been mutated. These results demonstrate that APC(Ama1) functions to coordinate the exit from meiosis II with cytokinesis.

      DOI: 10.1091/mbc.E08-06-0615

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    • Septins localize to microtubules during nutritional limitation in Saccharomyces cerevisiae 査読有り

      M. Evangelina Pablo-Hernando, Yolanda Arnaiz-Pita, Hiroyuki Tachikawa, Francisco del Rey, Aaron M. Neiman, Carlos R. Vazquez de Aldana

      BMC CELL BIOLOGY9   55   2008年10月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMED CENTRAL LTD  

      Background: In Saccharomyces cerevisiae, nutrient limitation stimulates diploid cells to undergo DNA replication and meiosis, followed by the formation of four haploid spores. Septins are a family of proteins that assemble a ring structure at the mother-daughter neck during vegetative growth, where they control cytokinesis. In sporulating cells, the septin ring disassembles and septins relocalize to the prospore membrane.
      Results: Here, we demonstrate that nutrient limitation triggers a change in the localization of at least two vegetative septins (Cdc10 and Cdc11) from the bud neck to the microtubules. The association of Cdc10 and Cdc11 with microtubules persists into meiosis, and they are found associated with the meiotic spindle until the end of meiosis II. In addition, the meiosis-specific septin Spr28 displays similar behavior, suggesting that this is a common feature of septins. Septin association to microtubules is a consequence of the nutrient limitation signal, since it is also observed when haploid cells are incubated in sporulation medium and when haploid or diploid cells are grown in medium containing non-fermentable carbon sources. Moreover, during meiosis II, when the nascent prospore membrane is formed, septins moved from the microtubules to this membrane. Proper organization of the septins on the membrane requires the sporulation-specific septins Spr3 and Spr28.
      Conclusion: Nutrient limitation in S. cerevisiae triggers the sporulation process, but it also induces the disassembly of the septin bud neck ring and relocalization of the septin subunits to the nucleus. Septins remain associated with microtubules during the meiotic divisions and later, during spore morphogenesis, they are detected associated to the nascent prospore membranes surrounding each nuclear lobe. Septin association to microtubules also occurs during growth in non-fermentable carbon sources.

      DOI: 10.1186/1471-2121-9-55

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    • Association of human DNA helicase RecQ5 beta with RNA polymerase II and its possible role in transcription 査読有り

      Keiichi Izumikawa, Mitsuaki Yanagida, Toshiya Hayano, Hiroyuki Tachikawa, Wataru Komatsu, Akira Shimamoto, Kazunobu Futami, Yasuhiro Furuichi, Takashi Shinkawa, Yoshio Yamauchi, Toshiaki Isobe, Nobuhiro Takahashi

      BIOCHEMICAL JOURNAL413 ( 3 ) 505 - 516   2008年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PORTLAND PRESS LTD  

      Although RecQ5 beta is a ssDNA (single-stranded DNA)-stimulated ATPase and an ATP-dependent DNA helicase with strand-annealing activities, its cellular function remains to be explored. In the present paper, we used immunopurification and MS-based analyses to show that human DNA helicase RecQ5 beta is associated with at least four RNAP II (RNA polymerase II) subunits. RecQ5 beta was also present in complexes immunoprecipitated using three different antibodies against the large subunit of RNAP II, or in complexes immunoprecipitated using an anti-FLAG antibody against either FLAG-RNAP II 33 kDa subunit or FLAG-Pin 1. Different regions of the non-helicase domain of the RecQ5 beta molecule were associated with hypophosphorylated and hyperphosphorylated forms of the RNAP II large subunit independently of DNA and RNA. RecQ5 beta was also found in nuclear chromatin fractions and associated with the coding regions of the LDL (low-density lipoprotem) receptor and beta-actin genes. Knockdown of the RecQ5 beta transcript increased the transcription of those genes. The results of the present study suggest that RecQ5 beta has suppressive roles in events associated with RNAP II-dependent transcription.

      DOI: 10.1042/BJ20071392

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    • Schizosaccharomyces pombe Sst4p, a conserved Vps27/Hrs homolog, functions downstream of phosphatidylinositol 3-kinase Pik3p to mediate proper spore formation 査読有り

      Masayuki Onishi, Michihiro Iida, Takako Koga, Sadayuki Yamada, Aiko Hirata, Tomoko Iwaki, Kaoru Takegawa, Yasuhisa Fukui, Hiroyuki Tachikawa

      EUKARYOTIC CELL6 ( 12 ) 2343 - 2353   2007年12月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

      Sporulation of the fission yeast Schizosaccharomyces pombe is a developmental process that generates gametes and that includes the formation of spore envelope precursors called the forespore membranes. Assembly and development of forespore membranes require vesicular trafficking from other intracellular membrane compartments. We have shown that phosphatidylinositol 3-kinase (PtdIns 3-kinase) is required for efficient and proper development of forespore membranes. The role of a FYVE domain protein, Sst4p, a homolog of Vps27p/Hrs, as a downstream factor for PtdIns 3-kinase in sporulation was investigated. sst4 Delta asci formed spores with oval-shaped morphology and with reduced viability compared to that of the wild-type spores. The extension of forespore membranes was inefficient, and bubble-like structures emerged from the leading edges of the forespore membranes. Sst4p localization was examined using fluorescent protein fusions and was found to be adjacent to the forespore membranes during sporulation. The localization and function of Sst4p were dependent on its FYVE domain and on PtdIns 3-kinase. Sst4p colocalized and interacted with Hse1p, a homolog of Saccharomyces cerevisiae Hse1p and of mammalian STAM. Mutations in all three UIM domains of the Sst4p/Hse1p complex resulted in formation of spores with abnormal morphology. These results suggest that Sst4p is a downstream factor of PtdIns 3-kinase and functions in forespore membrane formation.

      DOI: 10.1128/EC.00211-07

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    • Pleckstrin-2 selectively interacts with phosphatidylinositol 3-kinase lipid products and regulates actin organization and cell spreading 査読有り

      Norihisa Hamaguchi, Sayoko Ihara, Tsutomu Ohdaira, Hiromichi Nagano, Akihiro Iwamatsu, Hiroyuki Tachikawa, Yasuhisa Fukui

      BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS361 ( 2 ) 270 - 275   2007年9月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

      Pleckstrin-2 (PLEK2) has been implicated to be regulated by phosphatidylinositol (PI) 3-kinase, while pleckstrin1 (PLEK1) has been suggested to be a major PKC substrate in platelets. In this paper, we confirmed that PLEK2 specifically bound to the PI 3-kinase products in vitro and explored its behavior. PLEK2 was found to be expressed in various adherent cell lines, while PLEK1 expression was restricted to non-adherent cells in the protein level. Expression of PLEK2 in COS1 cells induced formation of protrusive F-actin structure and enhanced the actin rearrangements induced on collagen- or fibronectin-coated plates. A PLEK2 mutant incapable of binding to the PI 3-kinase products did not show any effect on actin rearrangement. Knockdown of PLEK2 by shRNA inhibited spreading of HCC2998 adenocarcinoma cells. PLEK2 colocalized with Rac and was suggested to be oligomerized. These results suggest that PLEK2 is involved in actin rearrangement in a PI 3-kinase dependent manner. (c) 2007 Elsevier Inc. All rights reserved.

      DOI: 10.1016/j.bbrc.2007.06.132

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    • SWAP-70 is required for oncogenic transformation by v-Src in mouse embryo fibroblasts 査読有り

      Yasuhisa Fukui, Takayuki Tanaka, Hiroyuki Tachikawa, Sayoko Ihara

      BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS356 ( 2 ) 512 - 516   2007年5月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

      SWAP-70 is a phosphatidylinositol trisphosphate (PtdIns(3,4,5)P-3) binding protein, which acts in F-actin rearrangement. The role of SWAP-70 in oncogenic transformation of mouse embryo fibroblasts (MEFs) by v-Src was examined by use of MEFs defective in SWAP-70. v-Src morphologically transformed MEFs lacking SWAP-70, but growth of the transformed cells in culture was slower than that of cells supplemented with exogenous SWAP-70. The v-Src-transformed MEFs deficient in SWAP-70 were unable to grow in soft agar while those expressing SWAP70 readily formed colonies, suggesting that SWAP-70 is required for anchorage independent growth of v-Src transformed MEFs. When transplanted in nude mice, tumors formed by the v-Src transformed SWAP-70(-/-) MEFs were smaller than those formed by cells expressing exogenous SWAP-70. These results suggest that SWAP-70 may be required for oncogenic transformation and contributes to cell growth in MEFs transformed by v-Src. (c) 2007 Elsevier Inc. All rights reserved.

      DOI: 10.1016/j.bbrc.2007.03.011

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    • Muc4 is required for activation of ErbB2 in signet ring carcinoma cell lines 査読有り

      Atsushi Yokoyama, Bin-Hai Shi, Takayuki Kawai, Hiroaki Konishi, Ryota Andoh, Hiroyuki Tachikawa, Sayoko Ihara, Yasuhisa Fukui

      BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS355 ( 1 ) 200 - 203   2007年3月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

      Signet-ring cell carcinoma is one of the most malignant tumors, classified histologically as a poorly differentiated adenocarcinoma. The ErbB2/ErbB3 complex is often constitutively activated, which suggests that the ErbB2/ErbB3 signaling pathway may be important for malignancy of this tumor. However, the mechanism underlying this activation has not been understood. Here, we show that ErbB2 and Muc4 bind in signet ring carcinoma cells, which was not seen in highly differentiated adenocarcinorna cell lines. ErbB3 was suggested to be a substrate of ErbB2 because knockdown of ErbB2 resulted in less phosphorylation of ErbB3. Inhibition of expression of Muc4 at the cell surface by the treatment of the cells with benzyl-GaINac, an inhibitor of mucin secretion, blocked phosphorylation of ErbB3, suggesting that activity of Erb132 depends on the expression of Muc4. These results supply the biochemical backgrounds in recent studies suggesting the contribution of Muc4 in the tumorigenesis. (c) 2007 Elsevier Inc. All rights reserved.

      DOI: 10.1016/j.bbrc.2007.01.133

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    • Activity of beta 3-beta 4 loop of the PH domain is required for the membrane targeting of SWAP-70 査読有り

      Yasuhisa Fukui, Isamu Wakamatsu, Hiroyuki Tachikawa, Yoshihiko Okamura, Takayuki Tanaka, Sayoko Ihara

      IUBMB LIFE59 ( 2 ) 99 - 103   2007年2月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS INC  

      SWAP-70 translocates to the plasma membrane in a phosphoinositide 3-kinase (PI 3-kinase)-dependent manner and contributes to membrane ruffling. It binds to phosphatidylinositol trisphosphate (PtdIns(3,4,5)P-3) through its PH domain, which is essential for the membrane translocation after EGF stimulation. We examined the behavior of the SWAP-70s which have mutations in the beta 3/beta 4 loop of the PH domain. The two mutants fused to green fluorescent protein (GFP) carrying the mutations failed to translocate to the plasma membrane. The sole PH domains carrying the same mutations behaved similarly. The PtdIns(3,4,5)P-3 binding activity of two mutants was comparable to that of the wild-type protein. These results suggest that translocation of SWAP-70 largely depends on the activity of the PH domain, and that not only PtdIns(3,4,5)P-3 binding activity, but also some additional activity of the PH domain is required for the translocation.

      DOI: 10.1080/15216540701222880

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    • TRIM11 binds to and destabilizes a key component of the activator-mediated cofactor complex (ARC105) through the ubiquitin-proteasome system 査読有り

      Hideaki Ishikawa, Hiroyuki Tachikawa, Yutaka Miura, Nobuhiro Takahashi

      FEBS LETTERS580 ( 20 ) 4784 - 4792   2006年9月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

      TRIM11 is a member of the tripartite-motif-containing protein family and is known to destabilize humanin, an inhibitor of Alzheimer-like neuronal insults. In this study, we demonstrate that TRIM11 interacts with activator-recruited cofactor 105-kDa component (ARC105) that mediates chromatin-directed transcription activation and is a key regulatory factor for transforming growth factor P (TGFO) signaling. Coexpression of TRIM11 increased ARC105 degradation but a proteasome inhibitor suppressed this. Co-expression of TRIM11 and ARC105 also increased ubiquitination of ARC105. In addition, TRIM11 suppressed ARC105-mediated transcriptional activation induced with TGFO in a reporter assay. These results suggest that TRIM11, with the ubiquitin-proteasome pathway, regulates ARC105 function in TGF beta signaling. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

      DOI: 10.1016/j.febslet.2006.07.066

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    • Alg14 recruits alg13 to the cytoplasmic face of the endoplasmic reticulum to form a novel bipartite UDP-N-acetylglucosamine transferase required for the second step of N-linked glycosylation 査読有り

      XD Gao, H Tachikawa, T Sato, Y Jigami, N Dean

      JOURNAL OF BIOLOGICAL CHEMISTRY280 ( 43 ) 36254 - 36262   2005年10月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

      N-linked glycosylation requires the synthesis of an evolutionarily conserved lipid-linked oligosaccharide (LLO) precursor that is essential for glycoprotein folding and stability. Despite intense research, several of the enzymes required for LLO synthesis have not yet been identified. Here we show that two poorly characterized yeast proteins known to be required for the synthesis of the LLO precursor, GlcNAc(2)-PP-dolichol, interact to form an unusual hetero-oligomeric UDP-GlcNAc transferase. Alg13 contains a predicted catalytic domain, but lacks any membrane-spanning domains. Alg14 spans the membrane but lacks any sequences predicted to play a direct role in sugar catalysis. We show that Alg14 functions as a membrane anchor that recruits Alg13 to the cytosolic face of the ER, where catalysis of GlcNAc(2)-PP-dol occurs. Alg13 and Alg14 physically interact and under normal conditions, are associated with the ER membrane. Overexpression of Alg13 leads to its cytosolic partitioning, as does reduction of Alg14 levels. Concomitant Alg14 overproduction suppresses this cytosolic partitioning of Alg13, demonstrating that Alg14 is both necessary and sufficient for the ER localization of Alg13. Further evidence for the functional relevance of this interaction comes from our demonstration that the human ALG13 and ALG14 orthologues fail to pair with their yeast partners, but when co-expressed in yeast can functionally complement the loss of either ALG13 or ALG14. These results demonstrate that this novel UDP-GlcNAc transferase is a unique eukaryotic ER glycosyltransferase that is comprised of at least two functional polypeptides, one that functions in catalysis and the other as a membrane anchor.

      DOI: 10.1074/jbc.M507569200

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    • Calcineurin inhibitors block dorsal-side signaling that affect late-stage development of the heart, kidney, liver, gut and somitic tissue during Xenopus embryogenesis 査読有り

      Y Yoshida, S Kim, K Chiba, S Kawai, H Tachikawa, N Takahashi

      DEVELOPMENT GROWTH & DIFFERENTIATION46 ( 2 ) 139 - 152   2004年4月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING ASIA  

      Calcineurin, a calcium/calmodulin-dependent serine/threonine protein phosphatase, is a key constituent of signaling pathways involved in antigen-dependent T-cell activation and development of the mammalian heart. In addition, calcineurin constitutes a part of the Wnt/calcium-signaling pathway that regulates early stages of dorsoventral axis formation in Xenopus embryos. Although some of the Wnt family members are involved in organ formation at relatively late stages of Xenopus development, the involvement of calcineurin in the development of those organs remains unclear. In the present study, we demonstrate that calcineurin inhibitors (cyclosporine A, FK506, and FK520), but not non-calcineurin inhibitors (rapamycin and GPI1046) that bind the same intracellular receptor as that for FK506, induce edema and gut coiling disruption and exhibit teratogenesis in the kidney, heart, gut, liver, and somitic tissue during Xenopus development. The same effects were observed by injecting the calcineurin inhibitors into the dorsal side, but not ventral side, of blastomeres at the 4-cell stage, although the inhibitors did not affect dorsoventral axis formation. These results suggest that calcineurin is involved in dorsal-side signaling that leads to the formation of the heart, kidney, liver, gut and somitic tissue during Xenopus embryogenesis.

      DOI: 10.1111/j.1440-169X.2004.00733.x

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    • Saccharomyces cerevisiae QNS1 codes for NAD(+) synthetase that is functionally conserved in mammals 査読有り

      Y Suda, H Tachikawa, A Yokota, H Nakanishi, N Yamashita, Y Miura, N Takahashi

      YEAST20 ( 11 ) 995 - 1005   2003年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS LTD  

      NAD(+), an essential molecule involved in a variety of cellular processes, is synthesized through de novo and salvage pathways. NAD(+) synthetase catalyses the final step in both pathways. Here we show that this enzyme is encoded by the QNS1 gene in Saccharomyces cerevisiae. Expression of Escherichia coli or Bacillus subtilis NAD(+) synthetases was able to suppress the lethality of a qns1 deletion, while a B. subtilis NAD(+) synthetase mutant with lowered catalytic activity was not. Overexpression of QNS1 tagged with HA led to elevated levels of NAD(+) synthetase activity in yeast extracts, and this activity can be recovered by immunoprecipitation using anti-HA antibody. An allele of QNS1 was constructed that carries a point mutation predicted to reduce the catalytic activity. Overexpression of this allele, qns1(G521E), failed to elevate NAD(+) synthetase levels and qns1(G521E) could not rescue the lethality caused by the depletion of Qns1p. These results demonstrate that NAD(+) synthetase activity is essential for cell viability. A GFP-tagged version of Qns1p displayed a diffuse localization in both the nucleus and the cytosol. Finally, the rat homologue of QNS1 was cloned and shown to functionally replace yeast QNS1, indicating that NAD(+) synthetase is functionally conserved from bacteria to yeast and mammals. Copyright (C) 2003 John Wiley Sons, Ltd.

      DOI: 10.1002/yea.1008

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    • A Gip1p-Glc7p phosphatase complex regulates septin organization and spore wall formation 査読有り

      H Tachikawa, A Bloecher, K Tatchell, AM Neiman

      JOURNAL OF CELL BIOLOGY155 ( 5 ) 797 - 808   2001年11月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROCKEFELLER UNIV PRESS  

      Sporulation of Saccharomyces cerevisiae is a developmental process in which a single cell is converted into four haploid spores. GIP1, encoding a developmentally regulated protein phosphatase 1 interacting protein, is required for spore formation. Here we show that GIP1 and the protein phosphatase 1 encoded by GLC7 play essential roles in spore development. The gip1 Delta mutant undergoes meiosis and prospore membrane formation normally, but is specifically defective in spore wall synthesis. We demonstrate that in wild-type cells, distinct layers of the spore wall are deposited in a specific temporal order, and that gip1 Delta cells display a discrete arrest at the onset of spore wall deposition. Localization studies revealed that Gip1p and Glc7p colocalize with the septins in structures underlying the growing prospore membranes. Interestingly, in the gip1 Delta mutant, not only is Glc7p localization altered, but septins are also delocalized. Similar phenotypes were observed in a glc7-136 mutant, which expresses a Glc7p defective in interacting with Gip1p. These results indicate that a Gip1p-Glc7p phosphatase complex is required for proper septin organization and initiation of spore wall formation during sporulation.

      DOI: 10.1083/jcb.200107008

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    • Hut1 proteins identified in Saccharomyces cerevisiae and Schizosaccharomyces pombe are functional homologues involved in the protein-folding process at the endoplasmic reticulum 査読有り

      H Nakanishi, K Nakayama, A Yokota, H Tachikawa, N Takahashi, Y Jigami

      YEAST18 ( 6 ) 543 - 554   2001年4月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

      The Saccharomyces cerevisiae HUT1 gene (scHUT1) and the Schizosaccharomyces pombe hut1(+) gene (sphut1(+)) encode hydrophobic proteins with approximately 30% identity to a human UDP-galactose transporter-related gene (UGTrel1) product. These proteins show a significant similarity to the nucleotide sugar transporter and are conserved in many eukaryotic species, but their physiological functions are not known. Both scHUT1 and sphut1(+) genes are non-essential for cell growth under normal conditions, and their disruptants show no defects in the modification of O- and N-linked oligosaccharides, but are sensitive to a membrane-permeable reducing agent, dithiothreitol (DTT). Consistent with this phenotype, scHUT1 has genetic interaction with ERO1, which plays an essential role in the oxidation of secretory proteins at the endoplasmic reticulum (ER). Overexpression of the MPD1 or MPD2 genes, which were isolated as multicopy suppressors of protein disulphide isomerase (PDI) depletion, could not replace the essential function of PDI in Delta hut1 S. cerevisiae cells. Our results indicate that scHut1p and spHut1p are functional homologues, and their physiological function is to maintain the optimal environment for the folding of secretory pathway proteins in the ER. Copyright (C) 2001 John Wiley & Sons, Ltd.

      DOI: 10.1002/yea.707

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    • Characterization of the subpopulations of rat ascites hepatoma AH109A cells with different invasive and metastatic activities 査読有り

      Y Miura, M Miyauchi, K Kubota, Q Li, H Tachikawa, D Fujimoto, K Yagasaki

      ANIMAL CELL TECHNOLOGY: BASIC & APPLIED ASPECTS, VOL 10   205 - 209   1999年

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      記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER  

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    • Molecular cloning and characterization of mouse ficolin-A 査読有り

      Y Fujimori, S Harumiya, Y Fukumoto, Y Miura, K Yagasaki, H Tachikawa, D Fujimoto

      BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS244 ( 3 ) 796 - 800   1998年3月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC  

      A novel ficolin-related gene was isolated from the mouse liver lambda ZAPII cDNA library. The protein encoded by this gene consists of both collagen-and fibrinogen-like domains, which are common features of the ficolin family, and was named mouse ficolin-A. The amino acid sequence of mouse ficolin-A is 60.2, 59.8, 59.8, and 59.6% identical to those of porcine ficolin-alpha, -beta, human ficolin-1, and EBP-37/P35, respectively. Northern blot analysis showed that mRNA of mouse ficolin-A is highly expressed in liver and spleen. Immunoblot analysis using an anti-mouse ficolin-A antiserum showed that mouse ficolin-A is a plasma protein with binding activity to elastin and GlcNAc. (C) 1998 Academic Press.

      DOI: 10.1006/bbrc.1998.8344

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    • Overproduction of Mpd2p suppresses the lethality of protein disulfide isomerase depletion in a CXXC sequence dependent manner 査読有り

      H Tachikawa, W Funahashi, Y Takeuchi, H Nakanishi, R Nishihara, S Katoh, XD Gao, T Mizunaga, D Fujimoto

      BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS239 ( 3 ) 710 - 714   1997年10月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

      The third multicopy suppressor gene of the PDI1 deletion from Saccharomyces cerevisiae, MPD2, was isolated and characterized, The MPD2 gene encodes a protein with a putative signal sequence, ER retention signal, and a disulfide isomerase active site Like sequence, The amino acid sequence around the active site like sequence is similar to the thioredoxin-like domains of PDI and PDI related proteins, although the similarity is comparatively low. A Delta-pdi1 strain overproducing Mpd2p showed slow growth and was sensitive to 1 mM dithiothreitol. Mpd2p can be detected in wild type cells and is a glycoprotein. Although the MPD2 gene was not essential for growth, overexpression of the gene partially restored the maturation defect of carboxypeptidase Y caused by the PDI1 deletion. Mutagenesis analysis revealed that Mpd2p can compensate for the loss of PDI with its CXXC sequence. (C) 1997 Academic Press.

      DOI: 10.1006/bbrc.1997.7426

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    • Similarity between galectin-5 and rkCBP17.5 査読有り

      E Kondo, S Yokote, R Miyamoto, Y Miyake, H Kagawa, S Harumiya, M Totsuka, H Tachikawa, S Kaminogawa, D Fujimoto

      BIOMEDICAL RESEARCH-TOKYO17 ( 6 ) 491 - 494   1996年12月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMED RES FOUND  

      We compared biochemically and immunohistochemically rkCBP17.5, a rat kidney beta-galactoside-binding protein (Mr 17,500), with galectin-5, a beta-galactoside-binding protein (Mr 18,000), initially isolated from the rat lung. The result suggests that rkCBP17.5 is highly similar to galectin-5.

      DOI: 10.2220/biomedres.17.491

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    • Characterization of ficolins as novel elastin-binding proteins and molecular cloning of human ficolin-1 査読有り

      S Harumiya, K Takeda, T Sugiura, Y Fukumoto, H Tachikawa, K Miyazono, D Fujimoto, H Ichijo

      JOURNAL OF BIOCHEMISTRY120 ( 4 ) 745 - 751   1996年10月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN BIOCHEMICAL SOC  

      A novel elastin-binding protein, EBP-37, was recently identified and purified from human plasma, Its partial amino acid sequences showed significant homology to porcine ficolins, which were originally purified from porcine uterus membranes as multimeric proteins with fibrinogen- and collagen-like domains. Here we report the presence of ficolins in an elastin-binding fraction of porcine plasma and the direct binding of recombinant porcine filcolin-alpha to elastin, In addition, a cDNA encoding a human counterpart of porcine ficolins that is composed of 319 amino acids and is different from EBP-37 was cloned and named human ficolin-1. Northern blotting of various human tissues revealed that human ficolin-1 mRNA is highly expressed in peripheral blood leukocytes. These data suggested that there are at least two kinds of ficolin-related proteins in both pig and human, and they may function as plasma proteins with elastin-binding activities.

      DOI: 10.1093/oxfordjournals.jbchem.a021474

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    • A novel β-galactoside-binding lectin in cultured murine lymphocytic leukemia cells. 査読有り

      Mai A, Jung S, Tachikawa H, Fujimoto D

      Journal of Biochemistry120 ( 3 ) 478 - 480   1996年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1093/oxfordjournals.jbchem.a021436

      Scopus

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    • ISOLATION AND CHARACTERIZATION OF A YEAST GENE, MPD1, THE OVEREXPRESSION OF WHICH SUPPRESSES INVIABILITY CAUSED BY PROTEIN DISULFIDE-ISOMERASE DEPLETION 査読有り

      H TACHIKAWA, Y TAKEUCHI, W FUNAHASHI, T MIURA, XD GAO, D FUJIMOTO, T MIZUNAGA, K ONODERA

      FEBS LETTERS369 ( 2-3 ) 212 - 216   1995年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

      MPD1, a yeast gene the overexpression of which suppresses the inviability caused by the loss of protein disulfide isomerase (PDI) was isolated and characterized. The MPD1 gene product retained a single disulfide isomerase active site sequence (APWCGHCK), an N-terminal putative signal sequence, and a C-terminal endoplasmic reticulum (ER) retention signal, and was a novel member of the PDI family, The gene product, identified in yeast extract, contained core size carbohydrates. MPD1 was not essential for growth, but overexpression of the gene suppressed the maturation defect of carboxypeptidase Y caused by PDI1 deletion, indicative of the related function to PDI in the yeast ER.

      DOI: 10.1016/0014-5793(95)00750-4

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    • EBP-37, a new elastin-binding protein in human plasma: structural similarity to ficolins, transforming growth factor-β1-binding proteins. 査読有り

      Harumiya S, Omori A, Sugiura T, Fukumoto Y, Tachikawa H, Fujimoto D

      Journal of Biochemistry117 ( 5 ) 1029 - 1035   1995年5月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1093/oxfordjournals.jbchem.a124802

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    • Characterization of natural IgG antibody with anti-β-galactosyl specificity. 査読有り

      Fujita H, Jung S, Mukainaka Y, Tachikawa H, Fujimoto D, Kosuge T, Shirai T, Takeiri A, Suzuki Y

      Biomedical Research15 ( 1 ) 17 - 25   1994年2月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.2220/biomedres.15.17

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    • MOLECULAR-STRUCTURE OF A YEAST GENE, PDI1, ENCODING PROTEIN DISULFIDE ISOMERASE THAT IS ESSENTIAL FOR CELL-GROWTH 査読有り

      H TACHIKAWA, T MIURA, Y KATAKURA, T MIZUNAGA

      JOURNAL OF BIOCHEMISTRY110 ( 2 ) 306 - 313   1991年8月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN BIOCHEMICAL SOC  

      A genomic DNA clone for protein disulfide isomerase (PDI) of Saccharomyces cerevisiae was isolated by hybridization with synthesized oligonucleotide probes based on a partial amino acid sequence of yeast PDI. The introduction of a multiple copy plasmid carrying this fragment into yeast caused a tenfold increase in PDI specific activity and in the amount of PDI antigen in the extract. The gene on this fragment was named PDI1. The nucleotide sequence of the gene predicts a polypeptide of 522 amino acids with about 30% identity to mammalian PDIs. The predicted amino acid sequence contains an N-terminal signal peptide-like sequence, the C-terminal putative endoplasmic reticulum retention signal of yeast (HDEL), and two putative active site sequences of PDI (WCGHCK). The predicted polypeptide is acidic and contains five putative glycosylation sites, consistent with the molecular properties of the purified yeast PDI [T. Mizunaga et al. (1990) J. Biochem. 108, 846-851]. The PDI1 gene was mapped on chromosome III. A gene disruption experiment revealed that the PDI1 gene is essential for cell growth.

      DOI: 10.1093/oxfordjournals.jbchem.a123576

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    • GLYCOSYLATION SITE BINDING-PROTEIN AND PROTEIN DISULFIDE ISOMERASE ARE IDENTICAL AND ESSENTIAL FOR CELL VIABILITY IN YEAST 査読有り

      M LAMANTIA, T MIURA, H TACHIKAWA, HA KAPLAN, WJ LENNARZ, T MIZUNAGA

      PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA88 ( 10 ) 4453 - 4457   1991年5月

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      記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

      Glycosylation site binding protein (GSBP) has been shown to be identical to protein disulfide isomerase (PDI; EC 5.3.4.1) in a variety of multicellular organisms. We have utilized immunological and biochemical techniques to determine if GSBP and PDI are identical in yeast. Antiserum prepared against yeast GSBP identified in microsomes by its ability to be labeled with a peptide photoaffinity probe was found to recognize PDI purified from yeast. Moreover, this purified yeast PDI was found to be specifically labeled by the photoaffinity probe originally used to identify GSBP in a variety of eukaryotes. On the basis of these observations, we conclude that yeast GSBP and PDI are the same protein. The structure of the yeast PDI gene revealed a product with sequence similarity to higher eukaryotic PDI/GSBP. Disruption of this gene in yeast resulted in a recessive lethal mutation, indicating that PDI/GSBP is required for cell viability.

      DOI: 10.1073/pnas.88.10.4453

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    MISC

    • Involvement of p32, fibrillarin, and Nop52 in pre-90S particle separation during human ribosome biogenesis

      H. Yoshikawa, W. Komatsu, T. Hayano, Y. Miura, K. Homma, K. Izumikawa, H. Ishikawa, N. Miyazawa, H. Tachikawa, Y. Yamauchi, T. Isobe, N. Takahashi

      MOLECULAR BIOLOGY OF THE CELL22   2011年

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      記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC CELL BIOLOGY  

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    • 出芽酵母の前胞子膜伸長に必要なPP1複合体の機能解析

      大平孝博, 田中貴之, 福地栄太, 須田恭之, 高橋哲夫, 伊原さよ子, 舘川宏之

      日本農芸化学会大会講演要旨集2010   2010年

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    • 出芽酵母胞子形成に必須なPP1ターゲティングサブユニットであるGip1の解析

      田中貴之, 福地栄太, 高橋哲夫, 伊原さよ子, 舘川宏之

      日本農芸化学会大会講演要旨集2009   2009年

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    • Regulation of the ARC105-mediated transcription by TRIM11

      H. Ishikawa, H. Tachikawa, Y. Miura, N. Takahashi

      FEBS JOURNAL274   218 - 218   2007年7月

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      記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:BLACKWELL PUBLISHING  

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    • Proteomic analysis of RecQ5 beta-associated protein complex that may constitute a part of transcriptional machinery

      K Izumikawa, M Yanagida, T Hayano, H Tachikawa, A Shimamoto, Y Furuichi, T Isobe, N Takahashi

      FASEB JOURNAL19 ( 4 ) A873 - A873   2005年3月

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      記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:FEDERATION AMER SOC EXP BIOL  

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    • Possible involvement of human parvulin, a peptidyl-prolyl isomerase, in early stages of ribosome biogenesis in multi-cellular organisms

      S Fujiyama, M Yanagida, T Hayano, Y Miura, H Tachikawa, T Isobe, N Takahashi

      FASEB JOURNAL17 ( 5 ) A1307 - A1307   2003年3月

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      記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:FEDERATION AMER SOC EXP BIOL  

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    • Characterization of EBP-37 and ficolins as novel elastin-binding proteins in human and porcine plasmas and molecular cloning of human ficolin-1

      S Harumiya, H Ichijo, K Takeda, T Sugiura, Y Fukumoto, H Tachikawa, K Miyazono, D Fujimoto

      MATRIX BIOLOGY16 ( 2 ) 71 - 71   1997年5月

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      記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:GUSTAV FISCHER VERLAG  

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    講演・口頭発表等

    • 出芽酵母が胞子の膜と壁をつくりだすメカニズムとその応用 招待有り

      舘川宏之

      第24回酵母合同シンポジウム  2023年11月16日 

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      開催年月日: 2023年11月16日 - 2023年11月17日

      記述言語:日本語   会議種別:口頭発表(招待・特別)  

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    • メンブレンコンタクトサイトの再配置による脂質輸送と膜伸長の分子機構解明

      舘川宏之, 須田恭之

      第3回オルガネラ・ゾーン研究会  2019年11月26日 

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      開催年月日: 2019年11月26日 - 2019年11月27日

      会議種別:シンポジウム・ワークショップ パネル(公募)  

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    • 酵母の前胞子膜形成におけるリン脂質とオルガネラ接触部位の役割

      舘川 宏之

      日本農芸化学会2019年東京大会  2019年3月26日 

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      記述言語:日本語   会議種別:シンポジウム・ワークショップ パネル(指名)  

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    • 出芽酵母前胞子膜形成の研究−新しい膜構造を構築する分子メカニズムの理解を目指して− 招待有り

      舘川 宏之

      酵母細胞研究会  2018年7月13日 

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      記述言語:日本語   会議種別:口頭発表(招待・特別)  

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    担当経験のある科目(授業)

    • 2023年2月 
      微生物共生作用学特論 ( 東京農業大学 )

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    •  
      応用生命科学特論 ( 東京農工大学 )

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    •  
      細胞機能調節学 ( 宇都宮大学 )

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    •  
      細胞調節生化学 ( 東京大学 )

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    •  
      微生物バイオテクノロジー ( 東京大学 )

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    •  
      化学と生物 基礎 ( 東京大学 )

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    •  
      基礎生物化学 ( 東京大学 )

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    •  
      代謝生化学 ( 東京大学 )

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    •  
      細胞生物学 ( 東京大学 )

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    •  
      生物化学 ( 東京大学 )

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    所属学協会

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    共同研究・競争的資金等の研究

    • 出芽酵母前胞子膜への膜脂質供給の分子機構解明

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

      詳細を見る

      2023年4月 - 2026年3月

      課題番号:23K05006

      配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

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    • 出芽酵母前胞子膜形態形成の分子機構解明とその応用

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

      詳細を見る

      2020年4月 - 2023年3月

      課題番号:20K05782

      配分額:4290000円 ( 直接経費:3300000円 、 間接経費:990000円 )

      PP1複合体とSSV複合体の前胞子膜伸長における役割を明らかにするため以下の実験を行った。
      1 PP1複合体について、野生株とgip1破壊株でリン酸化ペプチドを濃縮してMS解析を行った。関係するタンパク質が複数得られたが、3連で行ったところ再現性よく違いの見られるペプチドが得られず、さらなる工夫と解析が必要であることがわかった。
      2 昨年度、Spo71のPxPモチーフを含むドメインと前胞子膜マーカーの融合タンパク質が、Vps13を前胞子膜にリクルートできるが、それだけではspo71破壊株の胞子形成を回復できないことを示し、Spo71にはVps13を前胞子膜にリクルートする以外にも役割があることを示した。そこで、この融合タンパク質に加えて、前胞子膜のPI4Pレベルを低下させるタンパク質を同時に発現させる実験を行い、spo71破壊株の胞子形成が部分的に回復することを明らかにした。これにより、Spo71はVps13を前胞子膜にリクルートするのに加えて、前胞子膜上のPI4Pのレベルの制御に関与することが示された。
      3 SSV複合体とともに働くと考えられるtetherタンパク質(Ist2, Scs2, Scs22, Tcb1-3)の遺伝子について、多重破壊株を作製しその表現型を調べた。6重破壊株は胞子形成をしたが、前胞子膜の形態、そして胞子の形態が異常になることを明らかにした。このことは、tetherタンパク質が前胞子膜の形態形成に関係することを示している。さらにICE2を加えて7重破壊株を作製したが、不安定な表現型を示したので、再度検討の必要がある。

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    • 出芽酵母前胞子膜伸長の分子機構解明-タンパク質脱リン酸化とリン脂質代謝制御-

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

      詳細を見る

      2017年4月 - 2020年3月

      課題番号:17K07712

      配分額:4810000円 ( 直接経費:3700000円 、 間接経費:1110000円 )

      出芽酵母の胞子形成は、ダイナミックな細胞内再編成の過程である。本研究では、この過程で出現する前胞子膜の形成機構を解析することにより、細胞内新規膜構造形成機構の解明を目指した。この過程において働く、1型脱リン酸化酵素については、そのターゲットに迫った。また、この過程に必須なVps13を含む複合体については、他のコンタクトサイトタンパク質と協調することにより、小胞体と前胞子膜のコンタクトサイトを形成して、リン脂質のレベルの制御と前胞子膜の伸長において働くことを明らかにした。Vps13は様々なコンタクトサイトに局在を変えるタンパク質として注目されており、その膜形成における働きの理解に貢献した。

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    • 出芽酵母の前胞子膜形成におけるリン脂質代謝制御機構とその役割の解明

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

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      2013年4月 - 2016年3月

      課題番号:25450094

      配分額:5200000円 ( 直接経費:4000000円 、 間接経費:1200000円 )

      出芽酵母の前胞子膜形成をモデルに、生体膜の形態形成の分子機構に関する研究を行った。前胞子膜の適切な伸長に必要な遺伝子の遺伝学的解析から、膜のリン脂質の一つであるPI4Pの合成酵素の複合体がこの過程に関与することが示された。また、PI4Pとその合成酵素、そしてPS, PAが前胞子膜に局在することが示され、さらに、PI4P合成酵素複合体の形成阻害、PI4P合成酵素の活性を持つサブユニットの分解、そして前胞子膜上のPI4Pの減少が、前胞子膜伸長に重要であることが示唆された。これらのことは、前胞子膜の伸長においてPI4Pの調節が重要であることを示し、生体膜形態形成の分子機構の一端が明らかになった。

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    • 出芽酵母前胞子膜形成の分子機構解明とその応用

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

      詳細を見る

      2010年 - 2012年

      課題番号:22580076

      配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

      出芽酵母の前胞子膜形成は生体膜形成のモデルとして研究されている。本研究では、前胞子膜伸長において働く1型プロテインホスファターゼのターゲティングサブユニットであるGip1のドメイン構造を明らかにし、局在化に必要なドメインと機能に必要なドメインが異なることを示した。また、前胞子膜形成に必須な膜タンパク質(Spo71,Spo73,Vps13)について解析を行い、Spo73の局在化に必要な配列を明らかにし、Spo71とSpo73が相互作用して働くことを示した。また、Spo73は、筋ジストロフィーと関係する機能未知のジスフェリンドメインのみからなっており、疾患の分子機構の解明に役立つ知見を得ることができた。

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    • プロリンペプチド異性化翻訳後修飾酵素の基質蛋白質フォーカスドプロテオミクス解析

      日本学術振興会  科学研究費助成事業 

      高橋 信弘, 舘川 宏之

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      2003年 - 2005年

      課題番号:15310139

      配分額:13300000円 ( 直接経費:13300000円 )

      本研究は、プロリンペプチド結合の異性化反応を触媒するペプチジルプロリルシストランスイソメラーゼ(PPIase)の基質タンパク質をプロテオミクスの手法を適用して網羅的に解析することを目的とした。そのために、ヒトのシクロフィリンイソタイプ9種類、FKBPイソタイプ11種類、parvulinのイソタイプ3種類の合計23種類及び他の種のPPIaseを対象として解析を行った。まず、シクロフィリン・FKBPのアイソフォーム遺伝子のクローニングとエピトープタグを融合させた形での発現ベクターの作製、ほ乳類培養細胞・Xenopus卵での発現、各種アフィニティー精製、MALDI-TOFMS・ESI-LC-MS/MS質量分析法及びそのデータ処理の各実験方法の最適化について研究した。これによって、各種細胞での発現及びより高感度で迅速な解析が可能となったので、それぞれのPPIaseの結合蛋白質の同定及びPPIase阻害剤を用いた解析を行った。その結果、約200種類が同定され、蛋白質代謝・細胞周期・細胞増殖及び維持・転写・ストレス応答・蛋白質分解・リボソーム合成・DNA代謝・RNAプロセッシング・蛋白質運搬・物質運搬・蛋白質生合成などに関わるタンパク質を含むPPIaseの機能ネットワークの概容が明らかになった。これらの中で、CyPGがRNAスプライシングに関わる一群の蛋白質に作用していること、また、FKBP25及びparvulin14がリボソーム生合成に関わる多くの蛋白質に作用しており、特に、parvulin14の場合、これがリボソーム合成の初期段階に関与すると考えられるトランス因子のリボソーム前駆体上へのリクルートに関与する可能性があることなど新しい知見を得た。

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    • 酵母の分泌タンパク質高次構造形成過程に関与する因子の解析

      日本学術振興会  科学研究費助成事業 

      舘川 宏之

      詳細を見る

      1996年 - 1996年

      課題番号:08760071

      配分額:1000000円 ( 直接経費:1000000円 )

      我々は、これまで酵母PDIに関する研究を行い、PDIが生育に必須であることを示した。また、ガラクトース依存性PDI変異株を作製し、PDIが分泌タンパク質の立体構造形成に重要な役割を果たしていることを示した。そして、作製した変異株の生育を多コピーで相補する遺伝子(マルチコピーサプレッサー遺伝子)のクローニングを行い、2つの新規遺伝子(MPD1,MPD2)を取得し、その構造を明らかにした。
      本研究では、Mpd1pとMpd2pの機能および関係を調べた。まず、グルタチオンS-トランスフェラーゼとの融合タンパク質として、大腸菌内で発現させることにより、Mpd1pがPDI活性を持つタンパク質であることを示した。Mpd2pは、融合タンパク質が不溶性となったため、今後検討が必要である。また、Mpd1pおよびMpd2pの活性中心様配列にin vitro mutagenesis法により変異を導入した結果、この領域がPDIの欠損のマルチコピーサプレッサーとして働く為には必須であることが明らかになった。これらより、Mpd1pとMpd2pはPDIと同様な機能を持つタンパク質であり、これらがPDIのかわりをすることによりPDIの欠損を相補することが示唆された。また、遺伝学的関係を知るため、MPD1,MPD2,EUG1遺伝子の三重破壊体酵母を作製したが、生育可能で、顕著な表現形は観察されなかった。
      今後は、PDI,Mpd1p,Mpd2p,Eug1pの基質特異性の違いや発現の違いを観察することにより、これらタンパク質の生体内における役割を明らかにしていく予定である。

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