2021/06/22 更新

写真b

ヒノ ミキ
日野 美紀
HINO Miki
*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*
理学部
職名*
助教
学位
博士(農学) ( 東京大学 )
連絡先
メールアドレス
研究キーワード
  • 糖鎖修飾

  • 自然免疫

  • GPIアンカー

  • オルガネラ

  • 学内職務経歴*
    • 2021年4月 - 現在 
      理学部   助教
    • 2017年4月 - 2021年3月 
      理学部   生命理学科   助教
     

    研究分野

    • ライフサイエンス / 細胞生物学

    • ライフサイエンス / 機能生物化学

    経歴

    • 2017年4月 - 現在 
      立教大学   理学部 生命理学科   助教

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    • 2012年4月 - 2017年3月 
      立教大学   理学部 生命理学科   研究員

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    学歴

    • 1984年4月 - 1989年3月 
      東京大学   農学部   農芸化学科

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      国名: 日本国

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    論文

    • Nuclear envelope localization of PIG-B is essential for GPI-anchor synthesis in Drosophila. 査読有り

      Yamamoto-Hino M, Katsumata E, Suzuki E, Maeda Y, Kinoshita T, Goto S

      Journal of Cell Science   2018年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1242/jcs.218024

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    • Spätzle-Processing Enzyme-independent Activation of the Toll Pathway in Drosophila Innate Immunity. 査読有り

      Yamamoto-Hino M, Goto S

      Cell Structure and Function41 ( 1 ) 55 - 60   2016年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1247/csf.16002

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    • Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing 査読有り

      Miki Yamamoto-Hino, Hideki Yoshida, Tomomi Ichimiya, Sho Sakamura, Megumi Maeda, Yoshinobu Kimura, Norihiko Sasaki, Kiyoko F. Aoki-Kinoshita, Akiko Kinoshita-Toyoda, Hidenao Toyoda, Ryu Ueda, Shoko Nishihara, Satoshi Goto

      GENES TO CELLS20 ( 6 ) 521 - 542   2015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:WILEY-BLACKWELL  

      Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, 1,4-galactosyltransferase I, 1,3-galactosyltransferase II, and 1,3-glucuronyltransferase. This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes.

      DOI: 10.1111/gtc.12246

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    • Dynamic regulation of innate immune responses in Drosophila by Senju-mediated glycosylation. 査読有り 国際誌

      Miki Yamamoto-Hino, Masatoshi Muraoka, Shu Kondo, Ryu Ueda, Hideyuki Okano, Satoshi Goto

      Proceedings of the National Academy of Sciences of the United States of America112 ( 18 ) 5809 - 14   2015年5月5日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      The innate immune system is the first line of defense encountered by invading pathogens. Delayed and/or inadequate innate immune responses can result in failure to combat pathogens, whereas excessive and/or inappropriate responses cause runaway inflammation. Therefore, immune responses are tightly regulated from initiation to resolution and are repressed during the steady state. It is well known that glycans presented on pathogens play important roles in pathogen recognition and the interactions between host molecules and microbes; however, the function of glycans of host organisms in innate immune responses is less well known. Here, we show that innate immune quiescence and strength of the immune response are controlled by host glycosylation involving a novel UDP-galactose transporter called Senju. In senju mutants, reduced expression of galactose-containing glycans resulted in hyperactivation of the Toll signaling pathway in the absence of immune challenges. Genetic epistasis and biochemical analyses revealed that Senju regulates the Toll signaling pathway at a step that converts Toll ligand Spatzle to its active form. Interestingly, Toll activation in immune-challenged wild type (WT) flies reduced the expression of galactose-containing glycans. Suppression of the degalactosylation by senju overexpression resulted in reduced induction of Toll-dependent expression of an antimicrobial peptide, Drosomycin, and increased susceptibility to infection with Gram-positive bacteria. These data suggest that Senju-mediated galactosylation suppresses undesirable Toll signaling activation during the steady state; however, Toll activation in response to infection leads to degalactosylation, which raises the immune response to an adequate level and contributes to the prompt elimination of pathogens.

      DOI: 10.1073/pnas.1424514112

      PubMed

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    • In Vivo RNAi-Based Screens: Studies in Model Organisms 査読有り

      Miki Yamamoto-Hino, Satoshi Goto

      GENES4 ( 4 ) 646 - 665   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)   出版者・発行元:MDPI AG  

      RNA interference (RNAi) is a technique widely used for gene silencing in organisms and cultured cells, and depends on sequence homology between double-stranded RNA (dsRNA) and target mRNA molecules. Numerous cell-based genome-wide screens have successfully identified novel genes involved in various biological processes, including signal transduction, cell viability/death, and cell morphology. However, cell-based screens cannot address cellular processes such as development, behavior, and immunity. Drosophila and Caenorhabditis elegans are two model organisms whose whole bodies and individual body parts have been subjected to RNAi-based genome-wide screening. Moreover, Drosophila RNAi allows the manipulation of gene function in a spatiotemporal manner when it is implemented using the Gal4/UAS system. Using this inducible RNAi technique, various large-scale screens have been performed in Drosophila, demonstrating that the method is straightforward and valuable. However, accumulated results reveal that the results of RNAi-based screens have relatively high levels of error, such as false positives and negatives. Here, we review in vivo RNAi screens in Drosophila and the methods that could be used to remove ambiguity from screening results.

      DOI: 10.3390/genes4040646

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    • Identification of Proteasome Components Required for Apical Localization of Chaoptin Using Functional Genomics 査読有り

      Hiroyuki Yano, Miki Yamamoto-Hino, Wakae Awano, Kiyoko F. Aoki-Kinoshita, Kayoko Tsuda-Sakurai, Hideyuki Okano, Satoshi Goto

      JOURNAL OF NEUROGENETICS26 ( 1 ) 53 - 63   2012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:INFORMA HEALTHCARE  

      The distinct localization of membrane proteins with regard to cell polarity is crucial for the structure and function of various organs in multicellular organisms. However, the molecules and mechanisms that regulate protein localization to particular subcellular domains are still largely unknown. To identify the genes involved in regulation of protein localization, the authors performed a large-scale screen using a Drosophila RNA interference (RNAi) library, by which Drosophila genes could be knocked down in a tissue-and stage-specific manner. Drosophila photoreceptor cells have a morphologically distinct apicobasal polarity, along which Chaoptin (Chp), a glycosylphosphatidylinositol (GPI)-anchored membrane protein, and the Na+, K+-ATPase are localized to the apical and basolateral domains, respectively. By examining the subcellular localization of these proteins, the authors identified 106 genes whose knockdown resulted in mislocalization of Chp and Na+, K+-ATPase. Gene ontology analysis revealed that the knockdown of proteasome components resulted in mislocalization of Chp to the basolateral plasma membrane. These results suggest that the proteasome is involved, directly or indirectly, in selective localization of Chp to the apical plasma membrane of Drosophila photoreceptor cells.

      DOI: 10.3109/01677063.2012.661497

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    • Cisterna-specific Localization of Glycosylation-related Proteins to the Golgi Apparatus 査読有り

      Miki Yamamoto-Hino, Masato Abe, Takako Shibano, Yuka Setoguchi, Wakae Awano, Ryu Ueda, Hideyuki Okano, Satoshi Goto

      CELL STRUCTURE AND FUNCTION37 ( 1 ) 55 - 63   2012年

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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 SOC CELL BIOLOGY  

      The Golgi apparatus is an intracellular organelle playing central roles in post-translational modification and in the secretion of membrane and secretory proteins. These proteins are synthesized in the endoplasmic reticulum (ER) and transported to the cis-, medial- and trans-cisternae of the Golgi. While trafficking through the Golgi, proteins are sequentially modified with glycan moieties by different glycosyltransferases. Therefore, it is important to analyze the glycosylation function of the Golgi at the level of cisternae. Markers widely used for cis-, medial-and trans-cisternae/trans Golgi network (TGN) in Drosophila are GM130, 120 kDa and Syntaxin16 (Syx16); however the anti-120 kDa antibody is no longer available. In the present study, Drosophila Golgi complex-localized glycoprotein-1 (dGLG1) was identified as an antigen recognized by the anti-120 kDa antibody. A monoclonal anti-dGLG1 antibody suitable for immunohistochemistry was raised in rat. Using these markers, the localization of glycosyltransferases and nucleotide-sugar transporters (NSTs) was studied at the cisternal level. Results showed that glycosyltransferases and NSTs involved in the same sugar modification are localized to the same cisternae. Furthermore, valuable functional information was obtained on the localization of novel NSTs with as yet incompletely characterized biochemical properties.

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    • Autophagy-Dependent Rhodopsin Degradation Prevents Retinal Degeneration in Drosophila 査読有り

      Ryosuke Midorikawa, Miki Yamamoto-Hino, Wakae Awano, Yoshimi Hinohara, Emiko Suzuki, Ryu Ueda, Satoshi Goto

      JOURNAL OF NEUROSCIENCE30 ( 32 ) 10703 - 10719   2010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:SOC NEUROSCIENCE  

      Recent studies have demonstrated protective roles for autophagy in various neurodegenerative disorders, including the polyglutamine diseases; however, the role of autophagy in retinal degeneration has remained unclear. Accumulation of activated rhodopsin in some Drosophila mutants leads to retinal degeneration, and although it is known that activated rhodopsin is degraded in endosomal pathways in normal photoreceptor cells, the contribution of autophagy to rhodopsin regulation has remained elusive. This study reveals that activated rhodopsin is degraded by autophagy in collaboration with endosomal pathways to prevent retinal degeneration. Light-dependent retinal degeneration in the Drosophila visual system is caused by the knockdown or mutation of autophagy-essential components, such as autophagy-related protein 7 and 8 (atg-7/atg-8), or genes essential for PE (phosphatidylethanolamine) biogenesis and autophagosome formation, including Phosphatidylserine decarboxylase (Psd) and CDP-ethanolamine:diacylglycerol ethanolaminephosphotransferase (Ept). The knockdown of atg-7/8 or Psd/Ept produced an increase in the amount of rhodopsin localized to Rab7, positive late endosomes. This rhodopsin accumulation, followed by retinal degeneration, was suppressed by overexpression of Rab7, which accelerated the endosomal degradation pathway. These results indicate a degree of cross talk between the autophagic and endosomal/lysosomal pathways. Importantly, a reduction in rhodopsin levels rescued Psd knockdown-induced retinal degeneration. Additionally, the Psd knockdown-induced retinal degeneration phenotype was enhanced by Ppt1 inactivation, which causes infantile neuronal ceroid lipofuscinosis, implying that autophagy plays a significant role in its pathogenesis. Collectively, the current data reveal that autophagy suppresses light-dependent retinal degeneration in collaboration with the endosomal degradation pathway and that rhodopsin is a key substrate for autophagic degradation in this context.

      DOI: 10.1523/JNEUROSCI.2061-10.2010

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    • Balanced ubiquitylation and deubiquitylation of Frizzled regulate cellular responsiveness to Wg/Wnt 査読有り

      Akiko Mukai, Miki Yamamoto-Hino, Wakae Awano, Wakako Watanabe, Masayuki Komada, Satoshi Goto

      EMBO JOURNAL29 ( 13 ) 2114 - 2125   2010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:NATURE PUBLISHING GROUP  

      Wingless (Wg)/Wnt has been proposed to exert various functions as a morphogen depending on the levels of its signalling. Therefore, not just the concentration of Wg/Wnt, but also the responsiveness of Wg/Wnt-target cells to the ligand, must have a crucial function in controlling cellular outputs. Here, we show that a balance of ubiquitylation and deubiquitylation of the Wg/Wnt receptor Frizzled determines the cellular responsiveness to Wg/Wnt both in mammalian cells and in Drosophila, and that the cell surface level of Frizzled is regulated by deubiquitylating enzyme UBPY/ubiquitin-specific protease 8 (USP8). Although ubiquitylated Frizzled underwent lysosomal trafficking and degradation, UBPY/USP8-dependent deubiquitylation led to recycling of Frizzled to the plasma membrane, thereby elevating its surface level. Importantly, a gain and loss of UBPY/USP8 function led to up-and down-regulation, respectively, of canonical Wg/Wnt signalling. These results unveil a novel mechanism that regulates the cellular responsiveness to Wg/Wnt by controlling the cell surface level of Frizzled. The EMBO Journal (2010) 29, 2114-2125. doi:10.1038/emboj.2010.100; Published online 21 May 2010

      DOI: 10.1038/emboj.2010.100

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    • Identification of genes required for neural-specific glycosylation using functional genomics. 査読有り

      Yamamoto-Hino M, Kanie Y, Awano W, Aoki-Kinoshita KF, Yano H, Nishihara S, Okano H, Ueda R, Kanie O, Goto S

      PLos Genetics6 ( 12 ) e1001254   2010年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1371/journal.pgen.1001254.

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    • Insight into the Regulation of Glycan Synthesis in Drosophila Chaoptin Based on Mass Spectrometry 査読有り

      Yoshimi Kanie, Miki Yamamoto-Hino, Yayoi Karino, Hiroki Yokozawa, Shoko Nishihara, Ryu Ueda, Satoshi Goto, Osamu Kanie

      PLOS ONE4 ( 5 ) e5434   2009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:PUBLIC LIBRARY SCIENCE  

      Background: A variety of N-glycans attached to protein are known to involve in many important biological functions. Endoplasmic reticulum (ER) and Golgi localized enzymes are responsible to this template-independent glycan synthesis resulting glycoforms at each asparagine residues. The regulation mechanism such glycan synthesis remains largely unknown.
      Methodology/Principal Findings: In order to investigate the relationship between glycan structure and protein conformation, we analyzed a glycoprotein of Drosophila melanogaster, chaoptin (Chp), which is localized in photoreceptor cells and is bound to the cell membrane via a glycosylphosphatidylinositol anchor. Detailed analysis based on mass spectrometry revealed the presence of 13 N-glycosylation sites and the composition of the glycoform at each site. The synthetic pathway of glycans was speculated from the observed glycan structures and the composition at each N-glycosylation site, where the presence of novel routes were suggested. The distribution of glycoforms on a Chp polypeptide suggested that various processing enzymes act on the exterior of Chp in the Golgi apparatus, although virtually no enzyme can gain access to the interior of the horseshoe-shaped scaffold, hence explaining the presence of longer glycans within the interior. Furthermore, analysis of Chp from a mutant (RNAi against dolichyl-phosphate a-D-mannosyltransferase), which affects N-glycan synthesis in the ER, revealed that truncated glycan structures were processed. As a result, the distribution of glycoforms was affected for the high-mannose-type glycans only, whereas other types of glycans remained similar to those observed in the control and wild-type.
      Conclusions/Significance: These results indicate that glycan processing depends largely on the backbone structure of the parent polypeptide. The information we obtained can be applied to other members of the LRR family of proteins.

      DOI: 10.1371/journal.pone.0005434

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    • Spatial and temporal regulation of glycosylation during Drosophila eye development 査読有り

      Hiroyuki Yano, Miki Yamamoto-Hino, Satoshi Goto

      CELL AND TISSUE RESEARCH336 ( 1 ) 137 - 147   2009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:SPRINGER  

      Glycosylation plays an essential role during development, in processes such as morphogen distribution, cell-to-cell communication, and extracellular matrix formation. Glycosylation is regulated during development in both a spatial and temporal manner. This study presents a detailed description of glycan distribution from late pupal to adult stages in Drosophila ommatidia by using nine different lectins. The lectins ConA, LCA, and DSA, which recognize high-mannose or complex types of N-linked glycans stain both apical and basolateral membranes of photoreceptor cells, whereas SBA, DBA, and ABA lectins, which bind to mucin-type O-glycans, label the inter-rhabdomeral space. The O-linked GlcNAc moiety is strongly labeled by WGA on the nuclear membrane. The localization of these glycans does not change throughout late pupal development. In contrast, the abundance of O-linked glycans, bisected oligosaccharides, and GlcNAc-containing glycans detected by PNA, PHA-E4, and WGA, respectively, is reduced in rhabdomeres and other subcellular domains during late pupal development. Some of these glycans have also been detected in the Golgi and/or putative secretory vesicles, suggesting their dynamic transport during development. These glycans, whose expression is dynamically regulated in a spatial and temporal manner, may therefore play critical roles in ommatidial development.

      DOI: 10.1007/s00441-009-0753-6

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    • N-glycosylation of the Drosophila neural protein Chaoptin is essential for its stability, cell surface transport and adhesive activity 査読有り

      Yu Hirai-Fujita, Miki Yamamoto-Hino, Osamu Kanie, Satoshi Goto

      FEBS LETTERS582 ( 17 ) 2572 - 2576   2008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:ELSEVIER SCIENCE BV  

      Glycosylation of proteins can modulate their function in a striking variety of systems, including immune responses, neuronal activities and development. The Drosophila protein, Chaoptin ( Chp), is essential for the development and maintenance of photoreceptor cells. This protein is heavily glycosylated, but the possible role of this glycosylation is not well- understood. Here we show that mutations introduced into about 1/ 3 of 16 potential N- linked glycosylation sites within Chp impaired its cell adhesive activities when expressed in Drosophila S2 cells. Mutation of 2/ 3 of the glycosylation sites resulted in a marked decrease in Chp protein abundance. These results suggest that Nlinked glycosylation of Chp is essential for its stability and activity. (c) 2008 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.

      DOI: 10.1016/j.febslet.2008.06.028

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    • Distinct functional units of the Golgi complex in Drosophila cells 査読有り

      H Yano, M Yamamoto-Hino, M Abe, R Kuwahara, S Haraguchi, Kusaka, I, W Awano, A Kinoshita-Toyoda, H Toyoda, S Goto

      PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA102 ( 38 ) 13467 - 13472   2005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:NATL ACAD SCIENCES  

      A striking variety of glycosylation occur in the Golgi complex in a protein-specific manner, but how this diversity and specificity are achieved remains unclear. Here we show that stacked fragments (units) of the Golgi complex dispersed in Drosophila imaginal disk cells are functionally diverse. The UDP-sugar transporter FRINGE-CONNECTION (FRC) is localized to a subset of the Golgi units distinct from those harboring SULFATELESS (SFL), which modifies glucosaminoglycans (GAGs), and from those harboring the protease RHOMBOID (RHO), which processes the glycoprotein SPITZ (SPI). Whereas the glycosylation and function of NOTCH are affected in imaginal disks of frc mutants, those of SPI and of GAG core proteins are not, even though FRC transports a broad range of glycosylation substrates, suggesting that Golgi units containing FRC and those containing SFL or RHO are functionally separable. Distinct Golgi units containing FRC and RHO in embryos could also be separated biochemically by immunoisolation techniques. We also show that Tn-antigen glycan is localized only in a subset of the Golgi units distributed basally in a polarized cell. We propose that the different localizations among distinct Golgi units of molecules involved in glycosylation underlie the diversity of glycan modification.

      DOI: 10.1073/pnas.0506681102

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    • A Green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling 査読有り

      S Karasawa, T Araki, M Yamamoto-Hino, A Miyawaki

      JOURNAL OF BIOLOGICAL CHEMISTRY278 ( 36 ) 34167 - 34171   2003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:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

      We have cloned a gene which encodes a fluorescent protein from the stony coral, Galaxeidae. This protein absorbs light maximally at 492 nm and emits green light at 505 nm, and as a result, we have designated it "Azami-Green (AG)." Despite sharing a similar spectral profile with enhanced green fluorescent protein (EGFP) ( Clontech), the most popular variant of the Aequorea victoria green fluorescent protein, the identity between these two proteins at the amino acid level is only 5.7%. However, since AG has a high extinction coefficient, fluorescence quantum yield, and acid stability, it produces brighter green fluorescence in cultured cells than EGFP. Similar to other fluorescent proteins isolated from coral animals, AG forms a tight tetrameric complex, resulting in poor labeling of subcellular structures such as the plasma membrane and mitochondria. We have converted tetrameric AG into a monomeric form by the introduction of three amino acid substitutions, which were recently reported to be effective for monomerizing the red fluorescent protein from Discosoma coral ( DsRed, Clontech). The resultant monomeric AG allowed for efficient fluorescent labeling of all of the subcellular structures and proteins tested while retaining nearly all of the brightness of the original tetrameric form. Thus, monomeric AG is a useful monomeric green-emitting fluorescent protein comparable to EGFP.

      DOI: 10.1074/jbc.M304063200

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    • An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein 査読有り

      R Ando, H Hama, M Yamamoto-Hino, H Mizuno, A Miyawaki

      PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA99 ( 20 ) 12651 - 12656   2002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:NATL ACAD SCIENCES  

      We have cloned a gene encoding a fluorescent protein from a stony coral, Trachyphyllia geoffroyi, which emits green, yellow, and red light. The protein, named Kaede, includes a tripeptide, His-Tyr-Gly, that acts as a green chromophore that can be converted to red. The red fluorescence is comparable in intensity to the green and is stable under usual aerobic conditions. We found that the green-red conversion is highly sensitive to irradiation with UV or violet light (350-400 nm), which excites the protonated form of the chromophore. The excitation lights used to elicit red and green fluorescence do not induce photoconversion. Under a conventional epifluorescence microscope, Kaede protein expressed in HeLa cells turned red in a graded fashion in response to UV illumination; maximal illumination resulted in a 2,000-fold increase in the ratio of red-to-green signal. These color-changing properties provide a simple and powerful technique for regional optical marking. A focused UV pulse creates an instantaneous plane source of red Kaede within the cytosol. The red spot spreads rapidly throughout the cytosol, indicating its free diffusibility in the compartment. The extensive diffusion allows us to delineate a single neuron in a dense culture, where processes originating from many different somata are present. Illumination of a focused UV pulse onto the soma of a Kaede-expressing neuron resulted in filling of all processes with red fluorescence, allowing visualization of contact sites between the red and green neurons of interest.

      DOI: 10.1073/pnas.202320599

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    • Apical vesicles bearing inositol 1,4,5-trisphosphate receptors in the Ca2+ initiation site of ductal epithelium of submandibular gland 査読有り

      M Yamamoto-Hino, A Miyawaki, A Segawa, E Adachi, S Yamashina, T Fujimoto, T Sugiyama, T Furuichi, M Hasegawa, K Mikoshiba

      JOURNAL OF CELL BIOLOGY141 ( 1 ) 135 - 142   1998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:ROCKEFELLER UNIV PRESS  

      In polarized epithelial cells, agonists trigger Ca2+ waves and oscillations. These patterns may be caused by the compartmentalization of inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ pools into specific regions. We have investigated the relationship between the distribution of IP3 receptors (IP(3)Rs) and the spatiotemporal pattern of Ca2+ signaling in the duct cells of the rat submandibular gland (SMG), Using immunofluorescence, although labeling was somewhat heterogeneous, the IP(3)Rs were colocalized to the apical pole of the duct cells. Immunoelectron microscopy identified small apical vesicles bearing IP(3)R2 in some types of duct cells. Real-time confocal imaging of intact ducts demonstrated that, after carbachol stimulation, an initial Ca2+ spike occurred in the apical region. Subsequently, repetitive Ca2+ spikes spread from the apical to the middle cytoplasm, These apical Ca2+ initiation sites were found only in some "pioneer cells," rather than in all duct cells. We performed both Ca2+ imaging and immunofluorescence on the same ducts and detected the strongest immunosignals of IP(3)R2 in the Ca2+ initiation sites of the pioneer cells. The subcellular localization and expression level of IP(3)Rs correlated strongly with the spatiotemporal nature of the intracellular Ca2+ signal and distinct Ca2+ responses among the rat SMG duct cells.

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    • Localization of inositol 1,4,5-trisphosphate receptors in the rat kidney 査読有り

      T Monkawa, M Hayashi, A Miyawaki, T Sugiyama, M Yamamoto-Hino, M Hasegawa, T Furuichi, K Mikoshiba, T Saruta

      KIDNEY INTERNATIONAL53 ( 2 ) 296 - 301   1998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:BLACKWELL SCIENCE INC  

      Inositol 1,4,5-trisphosphate (IP3) receptors (IP(3)Rs) serve as intracellular calcium release channels involved in signal transduction of various hormones in the kidney. Molecular cloning studies have shown that there are three types of IP3R, designated type 1, type 2, and type 3. To characterize their localizations in the rat kidney, we employed immunohistochemical studies using type-specific monoclonal antibodies that were raised against the 15 C-terminal amino acids of each type of IP3R. Type 1 was detected in glomerular mesangial cells and vascular smooth muscle cells. Type 2 was expressed exclusively in intercalated cells of collecting duels from the cortex to the inner medulla. Type 3 was expressed in vascular smooth muscle cells, glomerular mesangial cells, and some cells of cortical collecting ducts, probably principal cells. As to the subcellular distribution, type 1 and type 2 showed a homogenous distribution in the cytoplasm, whereas type 3 was present mainly in the basolateral portion of the cytoplasm. These results indicate that IP3R isoforms were expressed in a cell-specific manner. The heterogeneous subcellular localizations among the IP3R types suggests compartmentalization of distinct IP3-sensitive Ca2+ pools.

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    • Subtype-specific expression patterns of inositol 1,4,5-trisphosphate receptors in rat airway epithelial cells 査読有り

      T Sugiyama, M YamamotoHino, K Wasano, K Mikoshiba, M Hasegawa

      JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY44 ( 11 ) 1237 - 1242   1996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:HISTOCHEMICAL SOC INC  

      We investigated the immunohistochemical localization of inositol 1,4,5-trisphosphate receptor (IP(3)R) Types 1, 2, and 3 in rat airway epithelium using the monoclonal antibodies KM1112, KM1083, and KM1082 specific for each type of IP(3)R, The epithelium from trachea to distal intrapulmonary airways (bronchioles) showed positive immunoreactivity for all types of IP(3)R, However, cell type as well as subcellular site immunoreactivity for each type of IP(3)R varied, IP(3)R Type 1 was found only in the apical thin cytoplasmic area of ciliated cells throughout all airway levels. IP(3)R Type 2 was exclusively localized to the entire cytoplasm of ciliated cells from the trachea to bronchioles, IP(3)R Type 3 was expressed mainly in the supranuclear cytoplasm not only of ciliated cells at all airway levels but also in Clara cells of the bronchiolar epithelium. Double fluorescent staining using combinations of KM1083 and Wisteria floribunda lectin or anti-rat 10-KD Clara cell-specific protein antibody confirmed that the IP(3)R Type 2-positive cells were neither seromucous cells nor Clara cells, These results indicate that the expression of three types of IP(3)Rs in different cell types and subcellular sites may reflect diverse physiological functions of IP(3)Rs within airway epithelial cells, The double staining studies suggested that the anti-IP(3)R Type 2 monoclonal antibody KM1083 would be a specific cell marker for ciliated cells of the airway epithelium.

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    • Heterotetrameric complex formation of inositol 1,4,5-trisphosphate receptor subunits. 査読有り

      Monkawa T, Miyawaki A, Sugiyama T, Yoneshima H, Yamamoto-Hino M, Furuichi T, Saruta T, Hasegawa M, Mikoshiba K

      Journal of Biological Chemistry16 ( 270 ) 14700 - 14704   1995年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

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    • Immunohistochemical study of inositol 1,4,5-trisphosphate receptor type 3 in rat central nervous system. 査読有り

      Yamamoto-Hino M, Miyawaki A, Kawano H, Sugiyama T, Furuichi T, Hasegawa M, Mikoshiba K

      Neuroreport26 ( 6 ) 273 - 276   1995年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

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    • The calmodulin-binding domain in the mouse type 1 inositol 1,4,5-trisphosphate receptor. 査読有り

      Yamada M, Miyawaki A, Saito K, Nakajima T, Yamamoto-Hino M, Ryo Y, Furuichi T, Mikoshiba K

      Biochemical Journal15 ( 308 ) 83 - 88   1995年

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

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    • MONOCLONAL-ANTIBODIES DISTINCTIVELY RECOGNIZING THE SUBTYPES OF INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR - APPLICATION TO THE STUDIES ON INFLAMMATORY CELLS 査読有り

      T SUGIYAMA, A FURUYA, T MONKAWA, M YAMAMOTOHINO, S SATOH, K OHMORI, A MIYAWAKI, N HANAI, K MIKOSHIBA, M HASEGAWA

      FEBS LETTERS354 ( 2 ) 149 - 154   1994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:ELSEVIER SCIENCE BV  

      Monoclonal antibodies were raised that specifically recognize the COOH-terminal sequences and the loop sequences between the fifth and the sixth transmembrane spanning regions of human inositol 1,4,5-trisphosphate receptor (IP(3)R) type 1, 2 and 3. Western blot analysis using Jurkat cells, mouse cerebellum, COS-7 expressing IP(3)R type 3 cDNA showed that those monoclonal antibodies reacted specifically with each of these three IP(3)R subtypes and that they do not cross-react. These antibodies could be used for the specific immunoprecipitation of IP(3)Rs. Using these monoclonal antibodies, the expression profiles of IP(3)R-subtype proteins were found to be different among inflammatory cells such as macrophages, polymorphonuclear cells, mast cells, eosinophils, splenocytes, thymocytes and megakaryocytic cells. Usually, more than one type of IP(3)R were expressed in a cell. simultaneously. The observation of CMK cells under immunofluorescence confocal microscopy revealed that IP(3)R type 1 and type 2 are located at different subcellular fractions.

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    • SUBTYPES OF INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR IN HUMAN HEMATOPOIETIC-CELL LINES - DYNAMIC ASPECTS OF THEIR CELL-TYPE-SPECIFIC EXPRESSION 査読有り

      T SUGIYAMA, M YAMAMOTOHINO, A MIYAWAKI, T FURUICHI, K MIKOSHIBA, M HASEGAWA

      FEBS LETTERS349 ( 2 ) 191 - 196   1994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  

      Inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ signaling plays important roles in cellular responses to extracellular stimuli. We recently succeeded in cloning human counterparts of the three subtypes derived from separate genes. Using the cDNA sequences type-specific to these subtype receptors, we here analyzed the expression profile of IP(3)R subtypes in stimulated and unstimulated human hematopoietic cell lines representing T cells; B cells, neutrophils, macrophages, erythrocytes and megakaryocytes. Northern and dot blot analysis showed that each IP(3)R subtype is expressed differently in these cells and that the expression profile in each cell is dynamically changed upon stimuli which induce differentiation. Moreover, most of these cells were found to simultaneously express at least two different subtype receptors.

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    • CLONING AND CHARACTERIZATION OF HUMAN TYPE-2 AND TYPE-3 INOSITOL 1,4,5-TRISPHOSPHATE RECEPTORS 査読有り

      M YAMAMOTOHINO, T SUGIYAMA, K HIKICHI, MG MATTEI, K HASEGAWA, S SEKINE, K SAKURADA, A MIYAWAKI, T FURUICHI, M HASEGAWA, K MIKOSHIBA

      RECEPTORS & CHANNELS2 ( 1 ) 9 - 22   1994年

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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:HARWOOD ACAD PUBL GMBH  

      We have cloned cDNAs coding for human type 2 and type 3 and part of type 1 inositol 1,4,5-trisphosphate receptors (IP(3)Rs). The complete nucleotide sequences for type 2 and type 3 receptors were determined and the pharmacological properties of the latter were characterized. Human type 2 and type 3 IP(3)Rs are 2701 amino acids and 2671 amino acids long, respectively, and have significant sequence homologies as well as structural similarities including the six membrane-spanning regions near the C-termini when compared with the rat or mouse counterpart. COS-7 cells transfected with human type 3 IP(3)R showed characteristic inositol 1,4,5-trisphosphate (IP3)-binding properties with Kd values of 28.8 nM. The order of potency of competition with IP3 was Ins(1,4,5)P-3 (IP3) > Ins(2,4,5)P-3 > Ins(1,3,4,5)P-4 > Ins(1,2,3,4,5,6)P-6. Type 2 and type 3 IP(3)Rs were mapped to human chromosomes 12p11 and 6p21, respectively, by in situ hybridization. cDNA cloning of the human IP(3)Rs allowed us to identify the types of the receptor expressed in various human hematopoietic and lymphoma cell lines. The type 3 receptor was present in all of cell lines tested, while the type 1 or 2 receptor was expressed in only particular cell types. The diffential expression of the IP(3)R types could confer the cell-specific regulation on the IP3/Ca2+ signalling.

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      日本学術振興会  科学研究費助成事業 

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