2022/11/25 更新

写真b

ゴトウ サトシ
後藤 聡
GOTO Satoshi
*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 生命理学科
理学研究科 生命理学専攻 博士課程後期課程
理学研究科 生命理学専攻 博士課程前期課程
職名*
教授
学位
博士(理学) ( 東京大学 )
研究テーマ*
  • 私達のような多細胞生物は、多種多様な細胞が協調しあうことによって生存・活動することができる。このように重要な細胞間のやりとりは、細胞表面に提示または細胞外に分泌される蛋白質などを介して行われている。したがって、それらの蛋白質の機能やその制御を研究することは、多細胞生物を理解するうえで非常に重要である。私達は、そのような蛋白質がどのように調節されているかについて、特に翻訳後修飾という観点から研究を行っている。

  • 学内職務経歴*
    • 2012年4月 - 現在 
      理学部   生命理学科   教授
    • 2012年4月 - 現在 
      理学研究科   生命理学専攻 博士課程前期課程   教授
    • 2012年4月 - 現在 
      理学研究科   生命理学専攻 博士課程後期課程   教授
     

    研究分野

    • ライフサイエンス / 細胞生物学

    経歴

    • 2012年4月 - 現在 
      立教大学   理学研究科 生命理学専攻博士課程前期課程   教授

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    • 2012年4月 - 現在 
      立教大学   理学研究科 生命理学専攻博士課程後期課程   教授

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    • 2012年4月 - 現在 
      立教大学   理学部 生命理学科   教授

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    学歴

    • 1987年4月 - 1992年3月 
      東京大学   理学系研究科   生物化学専攻

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      国名: 日本国

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    • 1987年4月 - 1992年3月 
      東京大学   理学系研究科   生物化学専攻

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      国名: 日本国

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    • 1983年4月 - 1987年3月 
      東京大学   理学部   生物化学科

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      国名: 日本国

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    論文

    • SPPL3-dependent downregulation of the synthesis of (neo)lacto-series glycosphingolipid is required for the staining of cell surface CD59. 国際誌

      Kohei Kawaguchi, Miki Yamamoto-Hino, Satoshi Goto

      Biochemical and biophysical research communications571   81 - 87   2021年9月24日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      CD59 is a small glycoprotein modified with a glycophosphatidylinositol (GPI) anchor that prevents the formation of the membrane attack complex, thereby protecting host cells from lysis. A previous study identified that cell surface CD59 staining required the intramembrane protease signal peptide peptidase-like 3 (SPPL3). However, the effect of SPPL3 on the staining of CD59 remains unknown. This study shows that SPPL3 is essential for the surface labeling of CD59 but not of major GPI-anchored proteins. Surface CD59 staining requires the intramembrane protease activity of SPPL3 and SPPL3-mediated suppression of the (neo)lacto-series glycosphingolipids (nsGSLs)-but not N-glycan-synthesis pathway. The abundance of nsGSLs may affect complement-dependent cytotoxicity by altering the abundance or accessibility of cell surface CD59.

      DOI: 10.1016/j.bbrc.2021.06.093

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    • Hrd1-dependent Degradation of the Unassembled PIGK Subunit of the GPI Transamidase Complex.

      Kohei Kawaguchi, Miki Yamamoto-Hino, Yoshiko Murakami, Taroh Kinoshita, Satoshi Goto

      Cell structure and function46 ( 2 ) 65 - 71   2021年9月3日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      Glycosylphosphatidylinositol (GPI)-anchored proteins are post-transcriptionally modified with GPI and anchored to the plasma membrane. GPI is attached to nascent proteins in the endoplasmic reticulum by the GPI transamidase complex, which consists of PIGT, PIGK, GPAA1, PIGU, and PIGS. Of these, PIGK is a catalytic subunit that is unstable without PIGT. This study investigated the pathway by which unassembled PIGK not incorporated into the complex is degraded. We showed that unassembled PIGK was degraded via the proteasome-dependent pathway and that Hrd1 (also known as SYVN1), a ubiquitin ligase involved in the endoplasmic reticulum-associated degradation pathway, was responsible for degradation of unassembled PIGK.Key words: Glycosylphosphatidylinositol, GPI transamidase complex, protein stability, transamidation, ERAD.

      DOI: 10.1247/csf.21019

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    • Subunits of the GPI transamidase complex localize to the endoplasmic reticulum and nuclear envelope in Drosophila. 国際誌

      Kohei Kawaguchi, Miki Yamamoto-Hino, Nina Matsuyama, Emiko Suzuki, Satoshi Goto

      FEBS letters595 ( 7 ) 960 - 968   2021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)  

      A total of 10-20% of plasma membrane proteins are anchored by glycosylphosphatidylinositol (GPI). GPI is attached to proteins by GPI transamidase (GPI-T), which contains five subunits named PIGK, PIGS, PIGT, PIGU, and GPAA1. We previously reported that PIGT localizes near the nucleus in Drosophila. However, localizations of the other four subunits remain unknown. Here, we show that a catalytic subunit of GPI-T, PIGK, mainly localizes to the endoplasmic reticulum (ER), while the other four subunits localize to the nuclear envelope (NE) and ER. The NE/ER localization ratio of PIGS differs between cell types and developmental stages. Our results suggest that GPI-T catalyzes GPI attachment in the ER and the other four subunits may have other unknown functions in the NE.

      DOI: 10.1002/1873-3468.14048

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    • Role of the Drosophila YATA protein in the proper subcellular localization of COPI revealed by in vivo analysis.

      Maiko Saito, Minoru Nakayama, Kyota Fujita, Atsuko Uchida, Hiroyuki Yano, Satoshi Goto, Hitoshi Okazawa, Masaki Sone

      Genes & genetic systems95 ( 6 ) 303 - 314   2021年3月23日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      yata mutants of Drosophila melanogaster exhibit phenotypes including progressive brain shrinkage, developmental abnormalities and shortened lifespan, whereas in mammals, null mutations of the yata ortholog Scyl1 result in motor neuron degeneration. yata mutation also causes defects in the anterograde intracellular trafficking of a subset of proteins including APPL, which is the Drosophila ortholog of mammalian APP, a causative molecule in Alzheimer's disease. SCYL1 binds and regulates the function of coat protein complex I (COPI) in secretory vesicles. Here, we reveal a role for the Drosophila YATA protein in the proper localization of COPI. Immunohistochemical analyses performed using confocal microscopy and structured illumination microscopy showed that YATA colocalizes with COPI and GM130, a cis-Golgi marker. Analyses using transgenically expressed YATA with a modified N-terminal sequence revealed that the N-terminal portion of YATA is required for the proper subcellular localization of YATA. Analysis using transgenically expressed YATA proteins in which the C-terminal sequence was modified revealed a function for the C-terminal portion of YATA in the subcellular localization of COPI. Notably, when YATA was mislocalized, it also caused the mislocalization of COPI, indicating that YATA plays a role in directing COPI to the proper subcellular site. Moreover, when both YATA and COPI were mislocalized, the staining pattern of GM130 revealed Golgi with abnormal elongated shapes. Thus, our in vivo data indicate that YATA plays a role in the proper subcellular localization of COPI.

      DOI: 10.1266/ggs.20-00027

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    • Lamin is essential for nuclear localization of the GPI synthesis enzyme PIG-B and GPI-anchored protein production in Drosophila. 国際誌

      Miki Yamamoto-Hino, Kohei Kawaguchi, Masaya Ono, Kazuhiro Furukawa, Satoshi Goto

      Journal of cell science133 ( 6 )   2020年3月26日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      Membrane lipid biosynthesis is a complex process that occurs in various intracellular compartments. In Drosophila, phosphatidylinositol glycan-B (PIG-B), which catalyzes addition of the third mannose in glycosylphosphatidylinositol (GPI), localizes to the nuclear envelope (NE). Although this NE localization is essential for Drosophila development, the underlying molecular mechanism remains unknown. To elucidate this mechanism, we identified PIG-B-interacting proteins by performing immunoprecipitation followed by proteomic analysis. We then examined which of these proteins are required for the NE localization of PIG-B. Knockdown of Lamin Dm0, a B-type lamin, led to mislocalization of PIG-B from the NE to the endoplasmic reticulum. Lamin Dm0 associated with PIG-B at the inner nuclear membrane, a process that required the tail domain of Lamin Dm0. Furthermore, GPI moieties were distributed abnormally in the Lamin Dm0 mutant. These data indicate that Lamin Dm0 is involved in the NE localization of PIG-B and is required for proper GPI-anchor modification of proteins.

      DOI: 10.1242/jcs.238527

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    • Stability of the transamidase complex catalyzing GPI anchoring of proteins. 査読有り 国際誌

      Kawaguchi K, Sato T, Kondo S, Yamamoto-Hino M, Goto S

      Biochemical and biophysical research communications512 ( 3 ) 584 - 590   2019年5月7日

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      記述言語:英語   掲載種別:研究論文(学術雑誌)  

      DOI: 10.1016/j.bbrc.2019.03.103

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

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

      J. Cell Sci.131 ( 20 )   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 Struct. Funct.41 ( 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 国際誌

      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

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

      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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    • Structure, function and formation of glycans in Drosophila 査読有り

      Yamamoto-Hino M, Okano H, Kanie O, Goto S

      Glycans: Biochemistry, Characterization and Applications   165 - 188   2012年2月1日

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      Glycosylation of proteins shows two striking features: first, there are a large number of glycan structures that are involved in various biological events (diversity) and, second, certain structures are selectively added to specific sites within proteins (specificity) in a context-dependent manner. However, the mechanisms that regulate diversity and specificity are unknown. Over the last few decades, genetic studies in Drosophila have been used to examine the functions mediated by glycosylation in vivo. Recent improvements in the combined use of biochemical/biophysical, genetic, and cell biology approaches has allowed the rapid and detailed analysis of the structure, function and formation of glycans in Drosophila. This chapter aims to provide an overview of the different types of glycans and their biological function(s) in Drosophila. The regulation of glycan synthesis in Drosophila and mammals is also discussed. © 2012 by Nova Science Publishers, Inc. All rights reserved.

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      その他リンク: http://orcid.org/0000-0003-4147-6868

    • AP-1 clathrin adaptor and CG8538/Aftiphilin are involved in Notch signaling during eye development in Drosophila melanogaster 査読有り 国際誌

      Satoshi Kametaka, Ai Kametaka, Shinichi Yonekura, Mineyuki Haruta, Seiichi Takenoshita, Satoshi Goto, Satoshi Waguri

      JOURNAL OF CELL SCIENCE125 ( 3 ) 634 - 648   2012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:COMPANY OF BIOLOGISTS LTD  

      Clathrin adaptor protein complex-1 (AP-1) and its accessory proteins play a role in the sorting of integral membrane proteins at the trans-Golgi network and endosomes. Their physiological functions in complex organisms, however, are not fully understood. In this study, we found that CG8538p, an uncharacterized Drosophila protein, shares significant structural and functional characteristics with Aftiphilin, a mammalian AP-1 accessory protein. The Drosophila Aftiphilin was shown to interact directly with the ear domain of gamma-adaptin of Drosophila AP-1, but not with the GAE domain of Drosophila GGA. In S2 cells, Drosophila Aftiphilin and AP-1 formed a complex and colocalized at the Golgi compartment. Moreover, tissue-specific depletion of AP-1 or Aftiphilin in the developing eyes resulted in a disordered alignment of photoreceptor neurons in larval stage and roughened eyes with aberrant ommatidia in adult flies. Furthermore, AP-1-depleted photoreceptor neurons showed an intracellular accumulation of a Notch regulator, Scabrous, and downregulation of Notch by promoting its degradation in the lysosomes. These results suggest that AP-1 and Aftiphilin are cooperatively involved in the intracellular trafficking of Notch during eye development in Drosophila.

      DOI: 10.1242/jcs.090167

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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.

      DOI: 10.1247/csf.11037

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    • Multi-stage mass spectrometric information obtained by deconvolution of energy-resolved spectra acquired by triple-quadrupole mass spectrometry 査読有り

      Osamu Kanie, Yoshimi Kanie, Shusaku Daikoku, Yuki Shioiri, Ayako Kurimoto, Sachiko Mutsuga, Satoshi Goto, Yukishige Ito, Katsuhiko Suzuki

      RAPID COMMUNICATIONS IN MASS SPECTROMETRY25 ( 11 ) 1617 - 1624   2011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  

      Triple-quadrupole mass spectrometry (TQ-MS) provides the capability to carry out collision-induced dissociation (CID) and it offers advantages in quantification when connected with high-performance liquid chromatography through an electrospray ionization interface. However, although TQ-MS provides information on partial structures through the analysis of product ions obtained by CID experiments, the method only provides single-stage CID experiments, which limits the detailed structural information that can be obtained. Herein, a method of overcoming this limitation of TQ-MS is described. A spectrum obtained by energy-resolved mass spectrometry (ERMS) was used to deconvolute the fragmentation process, with a Galili-antigenic trisaccharide derivative being used as an example. A replot of the ERMS data showing the ratios of the product ions to the precursor ion resulted in a descriptive graph. Analysis of the sum of the ratios of individual product ions to the precursor ion at specific CID energies revealed that the members of a series of product ions were related to each other. The obtained relationships and the m/z values of the product ions provided information on the fragmentation process taking place during the dissociation, indicating that the ERMS spectrum obtained by TQ-MS contained equivalent information to that obtainable by multi-stage MS/MS (MS(n); n >= 2). This method may allow users of triple-quadrupole mass spectrometers to obtain MS(n)-type information by performing a single ERMS experiment, which is even advantageous over quadrupole ion trap (QIT)-MS/MS because CID experiments on individual first-generation product ions are not required. Copyright (C) 2011 John Wiley & Sons, Ltd.

      DOI: 10.1002/rcm.5031

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    • Fluorescence-monitored zero dead-volume nanoLC-microESI-QIT-TOF MS for analysis of fluorescently tagged glycosphingolipids 査読有り 国際誌

      Shusaku Daikoku, Yasunari Ono, Atsuko Ohtake, Yasuko Hasegawa, Eiichiro Fukusaki, Katsuhiko Suzuki, Yukishige Ito, Satoshi Goto, Osamu Kanie

      ANALYST136 ( 5 ) 1046 - 1050   2011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:ROYAL SOC CHEMISTRY  

      An analysis of the glycan processing event is of particular importance to understand the nontemplate dependent synthetic mechanism of the multiple glycosylation reactions taking place in the Golgi apparatus in connection with the post-translational modification of biomolecules. In our efforts to address the issue, we constructed an analysis platform using nano-liquid chromatography (LC), which also worked as a spray tip, with an optical-fiber-based blue (470 nm) light emitting diode (LED)-induced fluorescence (520 nm) detector coupled with a microelectrospray ionization (ESI)-quadrupole ion trap (QIT)-time of flight (TOF) mass spectrometer (MS). This system was designed to enable both quantitative and qualitative analyses of fluorescently tagged molecules such as BODIPY-tagged lactosylceramide. Owing to the zero dead volume after LC separation, an extremely high sensitivity was achieved for the quantitative analysis (260 amol). It was also shown that a simultaneous online structural analysis based on MS could be achieved for the same quantity of analyte. To further demonstrate its potential, an enzymatic reaction of fluorescently tagged lactosylceramide using sialyltransferase was carried out, and the conversion yield was obtained on the basis of fluorescence detection. In addition, the structural details of a product, sialyl lactosylceramide, were obtained by MS and MS/MS analyses.

      DOI: 10.1039/c0an00715c

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    • Identification of Genes Required for Neural-Specific Glycosylation Using Functional Genomics 査読有り 国際誌

      Miki Yamamoto-Hino, Yoshimi Kanie, Wakae Awano, Kiyoko F. Aoki-Kinoshita, Hiroyuki Yano, Shoko Nishihara, Hideyuki Okano, Ryu Ueda, Osamu Kanie, Satoshi Goto

      PLOS GENETICS6 ( 12 ) e1001254   2010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:PUBLIC LIBRARY SCIENCE  

      Glycosylation plays crucial regulatory roles in various biological processes such as development, immunity, and neural functions. For example, alpha 1,3-fucosylation, the addition of a fucose moiety abundant in Drosophila neural cells, is essential for neural development, function, and behavior. However, it remains largely unknown how neural-specific a1,3-fucosylation is regulated. In the present study, we searched for genes involved in the glycosylation of a neural-specific protein using a Drosophila RNAi library. We obtained 109 genes affecting glycosylation that clustered into nine functional groups. Among them, members of the RNA regulation group were enriched by a secondary screen that identified genes specifically regulating alpha 1,3-fucosylation. Further analyses revealed that an RNA-binding protein, second mitotic wave missing (Swm), upregulates expression of the neural-specific glycosyltransferase FucTA and facilitates its mRNA export from the nucleus. This first large-scale genetic screen for glycosylation-related genes has revealed novel regulation of fucTA mRNA in neural cells.

      DOI: 10.1371/journal.pgen.1001254

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

      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 国際誌

      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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    • Membrane Protein Location-Dependent Regulation by PI3K (III) and Rabenosyn-5 in Drosophila Wing Cells 査読有り 国際誌

      Masato Abe, Yuka Setoguchi, Tsubasa Tanaka, Wakae Awano, Kuniaki Takahashi, Ryu Ueda, Akira Nakamura, Satoshi Goto

      PLOS ONE4 ( 10 ) e7306   2009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:PUBLIC LIBRARY SCIENCE  

      The class III phosphatidylinositol-3 kinase (PI3K (III)) regulates intracellular vesicular transport at multiple steps through the production of phosphatidylinositol-3-phosphate (PI(3)P). While the localization of proteins at distinct membrane domains are likely regulated in different ways, the roles of PI3K (III) and its effectors have not been extensively investigated in a polarized cell during tissue development. In this study, we examined in vivo functions of PI3K (III) and its effector candidate Rabenosyn-5 (Rbsn-5) in Drosophila wing primordial cells, which are polarized along the apical-basal axis. Knockdown of the PI3K (III) subunit Vps15 resulted in an accumulation of the apical junctional proteins DE-cadherin and Flamingo and also the basal membrane protein beta-integrin in intracellular vesicles. By contrast, knockdown of PI3K (III) increased lateral membrane-localized Fasciclin III (Fas III). Importantly, loss-of-function mutation of Rbsn-5 recapitulated the aberrant localization phenotypes of beta-integrin and Fas III, but not those of DE-cadherin and Flamingo. These results suggest that PI3K (III) differentially regulates localization of proteins at distinct membrane domains and that Rbsn-5 mediates only a part of the PI3K (III)-dependent processes.

      DOI: 10.1371/journal.pone.0007306

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

      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 国際誌

      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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    • [Membrane traffic in Drosophila development].

      Masato Abe, Satoshi Goto

      Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme53 ( 16 Suppl ) 2193 - 9   2008年12月

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      記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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    • [Regulation of glycosylation in Golgi units].

      Satoshi Goto

      Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme53 ( 12 Suppl ) 1475 - 9   2008年9月

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      記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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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 国際誌

      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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    • Comparative RP-HPLC for rapid identification of glycopeptides and application in off-line LC-MALDI-MS analysis 査読有り 国際誌

      Yoshimi Kanie, Akiko Enomoto, Satoshi Goto, Osamu Kanie

      CARBOHYDRATE RESEARCH343 ( 4 ) 758 - 768   2008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:ELSEVIER SCI LTD  

      Despite the increasing attention being paid to the functions of glycoproteins, their structural analysis is still difficult and hinders functional investigations. Structural analysis of post-translationally modified proteins is thought to be achieved using methods frequently utilized in proteomics research; however, the same methods cannot be used for glycosylated proteins. One of the difficulties associated with the physiochemical properties of glycopeptides and peptides is that the detection of the former is considerably more difficult, because of the existence of glycoforms that increase molecular weight and reduces quantities of individual species. Thus, difficulties are often faced in finding glycopeptide(s) by using MS when analyzing peaks (or fractions) obtained after proteolytic digestion and HPLC. One simple yet difficult solution to this problem would be to develop a purification method that provides better resolution. Our intention has been to address this issue by using a combination of conventional methods. We found that a method consisting of a combination of rough fractionation using a reverse-phase cartridge column under acidic conditions and comparative RP-HPLC, where the two chromatograms obtained using phosphate and borate buffers under basic conditions were compared, is effective for MS-based structural analysis. The applicability of the method in glycoprotein analysis was examined using various samples including ribonuclease B (RNase B), IgG1, ovalbumin (OVA), and asialo fetuin (ASF). The results suggest that the method is useful in the analysis of glycoproteins. (c) 2007 Elsevier Ltd. All rights reserved.

      DOI: 10.1016/j.carres.2007.12.008

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    • Sequential enzymatic glycosyltransfer reactions on a microfluidic device: Synthesis of a glycosaminoglycan linkage region tetrasaccharide 査読有り 国際誌

      Yasunari Ono, Motohiro Kitajima, Shusaku Daikoku, Toshifumi Shiroya, Shoko Nishihara, Yoshimi Kanie, Katsuhiko Suzuki, Satoshi Goto, Osamu Kanie

      LAB ON A CHIP8 ( 12 ) 2168 - 2173   2008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:ROYAL SOC CHEMISTRY  

      A microfluidic chip carrying three reaction chambers was designed and constructed to examine sequential multiple enzymatic reactions. The synthesis of oligosaccharides in living cells is carried out in the Golgi apparatus where multiple enzymes such as glycosidase and glycosyltransferases act on a variety of substrates to generate glycoconjugates that include glycolipids and glycoproteins. The regulatory mechanism of the process however remains unknown. A microchip-based analysis platform may provide a valuable tool with which to address the issue by mimicking the Golgi function. We thus examined 3 sequential glycosyltransfer reactions on a chip, and succeeded in the synthesis of a tetrasaccharide using immobilized enzymes. Also, the kinetic parameters for a recently identified glycosyltransferase, proteoglycan GalT-I, were obtained for the first time.

      DOI: 10.1039/b809316d

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    • [Golgi units regulating diversity of glycosylation].

      Satoshi Goto

      Seikagaku. The Journal of Japanese Biochemical Society79 ( 1 ) 42 - 5   2007年1月

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      記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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

      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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    • Approach for functional analysis of glycan using RNA interference 査読有り 国際誌

      S Nishihara, R Ueda, S Goto, H Toyoda, H Ishida, M Nakamura

      GLYCOCONJUGATE JOURNAL21 ( 1-2 ) 63 - 68   2004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:KLUWER ACADEMIC PUBL  

      The elucidation of the biological role of glycan is one of the most important issues to be resolved following the genome project. RNA interference is becoming an efficient reverse genetic tool for studying gene function in model organisms, including C. elegans and Drosophila melanogaster. Our molecular evolutionary study has shown that a prototype of glycosyltransferases, which synthesize a variety of glycan structures in the Golgi apparatus, was conserved between mammals and Drosophila. For analyses of the basic physiological functions of glycans, we established the Drosophila inducible RNAi knockdown system and applied it to one glycosyltransferase and one transporter, proteoglycan UDP-galactose: beta-xylose 1,4galactosyltransferase I and the PAPS-transporter, respectively. If on the silencing of each gene induced ubiquitously under the control of a cytoplasmic actin promoter, the RNAi knockdown fly died, then the protein was indispensable for life. The expression of the target gene was disrupted specifically and the degree of interference was well correlated with the phenotype. The inducible RNAi knockdown fly obtained using the GAL4-UAS system will pave the way for the functional analysis of glycans.

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    • Molecular cloning and identification of 3 '- phosphoadenosine 5 '-phosphosulfate transporter 査読有り 国際誌

      S Kamiyama, T Suda, R Ueda, M Suzuki, R Okubo, N Kikuchi, Y Chiba, S Goto, H Toyoda, K Saigo, M Watanabe, H Narimatsu, Y Jigami, S Nishihara

      JOURNAL OF BIOLOGICAL CHEMISTRY278 ( 28 ) 25958 - 25963   2003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 BIOCHEMISTRY MOLECULAR BIOLOGY INC  

      Nucleotide sulfate, namely 3'-phosphoadenosine 5'-phosphosulfate (PAPS), is a universal sulfuryl donor for sulfation. Although a specific PAPS transporter is present in Golgi membrane, no study has reported the corresponding gene. We have identified a novel human gene encoding a PAPS transporter, which we have named PAPST1, and the Drosophila melanogaster ortholog, slalom (sll). The amino acid sequence of PAPST1 (432 amino acids) exhibited 48.1% identity with SLL (465 amino acids), and hydropathy analysis predicted the two to be type III transmembrane proteins. The transient expression of PAPST1 in SW480 cells showed a subcellular localization in Golgi membrane. The expression of PAPST1 and SLL in yeast Saccharomyces cerevisiae significantly increased the transport of PAPS into the Golgi membrane fraction. In human tissues, PAPST1 is highly expressed in the placenta and pancreas and present at lower levels in the colon and heart. An RNA interference fly of sll produced with a GAL4-UAS system revealed that the PAPS transporter is essential for viability. It is well known that mutations of some genes related to PAPS synthesis are responsible for human inherited disorders. Our findings provide insights into the significance of PAPS transport and post-translational sulfation.

      DOI: 10.1074/jbc.M302439200

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    • Distinct developmental modes and lesion-induced reactions of dendrites of two classes of Drosophila sensory neurons 査読有り 国際誌

      K Sugimura, M Yamamoto, R Niwa, D Satoh, S Goto, M Taniguchi, S Hayashi, T Uemura

      JOURNAL OF NEUROSCIENCE23 ( 9 ) 3752 - 3760   2003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:SOC NEUROSCIENCE  

      Little has been understood about the underlying mechanisms that generate the morphological diversity of dendritic trees. Dendritic arborization neurons in Drosophila provide an excellent model system to tackle this question, and they are classified into classes I-IV in order of increasing arbor complexity. Here we have developed transgenic green fluorescent protein markers for class I or class IV cells, which allowed time-lapse recordings of dendritic birth in the embryo, its maturation processes in the larva, and lesion-induced reactions. The two classes used distinct strategies of dendritic emergence from the cell body and branching, which contributed to differences in their basic arbor patterns. In contrast to the class I cells examined, one cell of class IV, which was a focus in this study, continued to elaborate branches throughout larval stages, and it was much more capable of responding to the severing of branches. We also investigated the cellular basis of field formation between adjacent class IV cells. Our results support the fact that class-specific inhibitory interaction is necessary and sufficient for tiling and confirmed that this intercellular communication was at work at individual dendrodendritic interfaces. Finally, this inhibitory signaling appeared to play a central role when arbors of adjacent cells started meeting midway between the cells and until the body wall became partitioned into abutting, minimal-overlapping territories.

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    • The role of Wg signaling in the patterning of embryonic leg primordium in Drosophila 査読有り 国際誌

      K Kubota, S Goto, S Hayashi

      DEVELOPMENTAL BIOLOGY257 ( 1 ) 117 - 126   2003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  

      Cellular interaction between the proximal and distal domains of the limb plays key roles in proximal-distal patterning. In Drosophila, these domains are established in the embryonic leg imaginal disc as a proximal domain expressing escargot, surrounding the Distal-less expressing distal domain in a circular pattern. The leg imaginal disc is derived from the limb primordium that also gives rise to the wing imaginal disc. We describe here essential roles of Wingless in patterning the leg imaginal disc. Firstly, Wingless signaling is essential for the recruitment of dorsal-proximal, distal, and ventral-proximal leg cells. Wingless requirement in the proximal leg domain appears to be unique to the embryo, since it was previously shown that Wingless signal transduction is not active in the proximal leg domain in larvae. Secondly, downregulation of Wingless signaling in wing disc is essential for its development, suggesting that Wg activity must be downregulated to separate wing and leg discs. In addition, we provide evidence that Dll restricts expression of a proximal leg-specific gene expression. We propose that those embryo-specific functions of Wingless signaling reflect its multiple roles in restricting competence of ectodermal cells to adopt the fate of thoracic appendages. (C) 2003 Elsevier Science (USA). All rights reserved.

      DOI: 10.1016/S0012-1606(03)00062-9

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    • GETDB, a database compiling expression patterns and molecular locations of a collection of Gal4 enhancer traps 査読有り

      S Hayashi, K Ito, Y Sado, M Taniguchi, A Akimoto, H Takeuchi, T Aigaki, F Matsuzaki, H Nakagoshi, T Tanimura, R Ueda, T Uemura, M Yoshihara, S Goto

      GENESIS34 ( 1-2 ) 58 - 61   2002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:WILEY-LISS  

      DOI: 10.1002/gene.10137

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    • UDP-sugar transporter implicated in glycosylation and processing of Notch 査読有り

      S Goto, M Taniguchi, M Muraoka, H Toyoda, Y Sado, M Kawakita, S Hayashi

      NATURE CELL BIOLOGY3 ( 9 ) 816 - 822   2001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:MACMILLAN PUBLISHERS LTD  

      Glycosylation modifies protein activities in various biological processes. Here, we report the functions of a novel UDP-sugar transporter (UST74C, an alternative name for Fringe connection (Frc)) localized to the Golgi apparatus in cellular signalling of Drosophila. Mutants in the frc gene exhibit phenotypes resembling wingless and Notch mutants. Both Fringe-dependent and Fringe-independent Notch pathways are affected, and both glycosylation and proteolytic maturation of Notch are defective in mutant larvae. The results suggest that changes in nucleotide-sugar levels can differently affect Wingless and two distinct aspects of Notch signalling.

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    • EGF receptor attenuates Dpp signaling and helps to distinguish the wing and leg cell fates in Drosophila 査読有り

      K Kubota, S Goto, K Eto, S Hayashi

      DEVELOPMENT127 ( 17 ) 3769 - 3776   2000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:COMPANY OF BIOLOGISTS LTD  

      Wing and leg precursors of Drosophila are recruited from a common pool of ectodermal cells expressing the homeobox gene Dll. Induction by Dpp promotes this cell fate decision toward the wing and proximal leg. We report here that the receptor tyrosine kinase EGFR antagonizes the wing-promoting function of Dpp and allows recruitment of leg precursor cells from uncommitted ectodermal cells. By monitoring the spatial distribution of cells responding to Dpp and EGFR, we show that nuclear transduction of the two signals peaks at different position along the dorsoventral axis when the fates of wing and leg discs are specified and that the balance of the two signals assessed within the nucleus determines the number of cells recruited to the wing. Differential activation of the two signals and the cross talk between them critically affect this cell fate choice.

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    • Proximal to distal cell communication in the Drosophila leg provides a basis for an intercalary mechanism of limb patterning 査読有り

      S Goto, S Hayashi

      DEVELOPMENT126 ( 15 ) 3407 - 3413   1999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:COMPANY OF BIOLOGISTS LTD  

      Proximodistal patterning in the Drosophila leg is elaborated from the circular arrangement of the proximal domain expressing escargot and homothorax, and the distal domain expressing Distal-less that are allocated during embryogenesis. The distal domain differentiates multiply segmented distal appendages by activating additional genes such as dachshund. Secreted signaling molecules Wingless and Decapentaplegic, expressed along the anterior-posterior compartment boundary, are required for activation of Distal-less and dachshund and repression of homothorax in the distal domain. However, whether Wingless and Decapentaplegic are sufficient for the circular pattern of gene expression is not known. Here we show that a proximal gene escargot and its activator homothorax ax regulate proximodistal patterning in the distal domain. Clones of cells expressing escargot or homothorax placed in the distal domain induce intercalary expression of dachshund in surrounding cells and reorient planar cell polarity of those cells. Escargot and homothorax-expressing cells also sort out from other cells in the distal domain. We suggest that inductive cell communication between the proximodistal domains, which is maintained in part by a cell-sorting mechanism, is the cellular basis for an intercalary mechanism of the proximodistal axis patterning of the limb.

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    • p38 mitogen-activated protein kinase can be involved in transforming growth factor- superfamily signal transduction in Drosophila wing morphogenesis. 査読有り

      Adachi-Yamada,T, Nakamura,M, Irie,K, Sano, Y Tomoyasu,Y, Sano,Y, Mori,E, Goto,S, Ueno,N, Nishida,Y, Matsumoto,K

      Mol. Cell. Biol19 ( 3 ) 2322 - 2329   1999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)  

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    • Cell migration within the embryonic limb primordium of Drosophila as revealed by a novel fluorescence method to visualize mRNA and protein 査読有り 国際誌

      S Goto, S Hayashi

      DEVELOPMENT GENES AND EVOLUTION207 ( 3 ) 194 - 198   1997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:SPRINGER VERLAG  

      We report a new technique using fluorescent probes to detect a mRNA and a protein simultaneously in the Drasophila embryo. For in situ hybridization, 3-hydroxy-N-2'-biphenyl-2-naphthalenecarboxamide phosphate ester (HNPP)/Fast Red TR was used as a fluorescent substrate for alkaline phosphatase. It was possible to compare protein and mRNA expression on a cell by cell basis with a laser scanning confocal microscope. We applied this technique to analyse the dynamics of Distal-less (Dll) enhancer activity in the thoracic limb primordium in the early Drosophila embryo. We stained embryos bearing the Dll early enhancer (Dll-304) fused to the Escherichia coli lacZ gene. LacZ mRNA was delectable in the ventral region of the limb primordium, and beta-galactosidase protein in the dorsal region. In the middle, both mRNA and protein were detectable. These results suggest that the Dll enhancer is activated in the ventral region of the Limb primordium and that Dll-positive cells migrate from a ventral position to a dorsal one within a single limb primordium.

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    • Specification of the embryonic limb primordium by graded activity of Decapentaplegic 査読有り

      S Goto, S Hayashi

      DEVELOPMENT124 ( 1 ) 125 - 132   1997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:COMPANY OF BIOLOGISTS LTD  

      Two thoracic limbs of Drosophila, the leg and the wing, originate from a common cluster of cells that include the source of two secreted signaling molecules, Decapentaplegic and Wingless. We show that Wingless, but not Decapentaplegic, is responsible for initial specification of the limb primordia with a distal identity. Limb formation is restricted to the lateral position of the embryo by negative control of the early function of Decapentaplegic and the EGF receptor homolog that determine the global dorsoventral pattern. Late function of Decapentaplegic locally determines two additional cell identities in a dosage dependent manner. Loss of Decapentaplegic activity results in a deletion of the proximal structures of the limb, which is in contrast to the consequence of decapentaplegic mutations in the imaginal disc, which cause a deletion of distal structures. The results indicate that the limb pattern elements are added in a distal to proximal direction in the embryo, which is opposite to what is happening in the growing imaginal disc. We propose that Wingless and Decapentaplegic act sequentially to initiate the proximodistal axis.

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    • Enhancer-trap detection of expression patterns corresponding to the polar coordinate system in the imaginal discs of Drosophila melanogaster 査読有り 国際誌

      Satoshi Goto, Teiichi Tanimura, Yoshiki Hotta

      Roux's Archives of Developmental Biology204 ( 6 ) 378 - 391   1995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-Verlag  

      We have isolated three classes of "enhancertrap" lines of Drosophila in which lacZ expression patterns in the imaginal discs are consistent with the idea of a polar (radial and angular) coordinate system of positional information. In the first class (HZ76), a circular pattern was expressed transiently during the mid-third instar larval stage when the radial components of the coordinate are probably generated. The expression patterns of the second class (HZ84) were sector-shaped and circular in the leg disc, suggesting a correlation with both radial and angular coordinate values. The expression patterns found in the third class (PZ63 and PZ22) were circular and appeared to reflect radial positional values. Expression in the latter two classes always began in the presumptive dorsal region of the leg disc and gradually spread to the ventral region. These developmental profiles of expression suggested the existence of a centre that initiates patterned gene expression in the presumptive dorsal region of the leg disc. The PZ22 line showed transient expression during tarsal segmentation, suggesting its involvement in tarsal segment formation. We have cloned the PZ22 gene and partially determined its sequence. The deduced amino acid sequence contained a zinc finger motif found in DNA/RNA binding proteins. By in situ hybridization, we determined that the PZ22 gene was transcribed in the leg disc in a pattern identical to that of the lacZ expression. In addition, it was expressed transiently in the embryonic mesoderm during mesoderm segmentation. The PZ22 gene, therefore, may function both in mesodermal segmentation in the embryo and in tarsal segmentation in the imaginal disc. © 1995 Springer-Verlag.

      DOI: 10.1007/BF00360483

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    • PURIFICATION, CHARACTERIZATION, AND AMINO-ACID-SEQUENCES OF PEPSINOGENS AND PEPSINS FROM THE ESOPHAGEAL MUCOSA OF BULLFROG (RANA-CATESBEIANA) 査読有り

      E YAKABE, M TANJI, M ICHINOSE, S GOTO, K MIKI, K KUROKAWA, H ITO, T KAGEYAMA, K TAKAHASHI

      JOURNAL OF BIOLOGICAL CHEMISTRY266 ( 33 ) 22436 - 22443   1991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:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

      Two pepsinogens (pepsinogens 1 and 2) were purified from the esophageal mucosa of the bullfrog (Rana catesbeiana), and their molecular weights were determined to be 40,100 and 39,200, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The NH2-terminal 70-residue sequences of both pepsinogens are the same, including the 36-residue activation segment. Furthermore, a cDNA clone encoding frog pepsinogen was obtained and sequenced, which permitted deduction of the complete amino acid sequence (368 residues) of one of the pepsinogen isozymogens. The calculated molecular weight of the protein (40,034) coincided well with the values obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results are incompatible with the previous report (Shugerman R. P., Hirschowitz, B. I., Bhown, A. S., Schrohenloher, R. E., and Spenney, J. G. (1982) J. Biol. Chem. 257, 795-798) that the major pepsinogen isolated from the bullfrog esophageal gland is a unique "mini" pepsinogen with a molecular weight of approximately 12,000-34,000.
      The two pepsinogens were immunologically indistinguishable from each other and related to human pepsinogen C. The deduced amino acid sequence was also more homologous with those of pepsinogens C than those of pepsinogens A and prochymosin. These results indicate that the frog pepsinogens belong to the pepsinogen C group. They were both glycoproteins, and therefore, this is the first finding of carbohydrate-containing pepsinogens C. Both pepsinogens were activated to pepsins in the same manner by an apparent one-step mechanism. The resulting pepsins were enzymatically indistinguishable from each other, and their properties resembled those of tuna pepsins.

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    • ISOLATION OF DROSOPHILA GENOMIC CLONES HOMOLOGOUS TO THE EEL SODIUM-CHANNEL GENE 査読有り

      H OKAMOTO, K SAKAI, S GOTO, E TAKASUISHIKAWA, Y HOTTA

      PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES63 ( 7 ) 284 - 288   1987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:JAPAN ACAD  

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    MISC

    • 翻訳後修飾と選別輸送をつかさどる小胞体・ゴルジ体の“オルガネラ・ゾーン” 招待有り 査読有り

      後藤聡, 山本(日野)美紀

      生体の科学69   551 - 555   2018年12月

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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    • In Vivo RNAi-Based Screens: Studies in Model Organisms 招待有り 査読有り 国際誌

      Miki Yamamoto-Hino, Satoshi Goto

      GENES4 ( 4 ) 646 - 665   2013年12月

      詳細を見る

      記述言語:英語   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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    • ユビキチン化によるFrizzledのリソソーム分解を介したWntシグナル強度の制御 査読有り

      Daocharad Burana, 後藤聡, 駒田雅之

      細胞工学32   396 - 400   2013年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    • Balanced ubiquitination determines cellular responsiveness to extracellular stimuli 招待有り 査読有り 国際誌

      Akiko Mukai, Miki Yamamoto-Hino, Masayuki Komada, Hideyuki Okano, Satoshi Goto

      CELLULAR AND MOLECULAR LIFE SCIENCES69 ( 23 ) 4007 - 4016   2012年12月

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      記述言語:英語   出版者・発行元:SPRINGER BASEL AG  

      Signal strength evoked by ligand stimulation is crucial for cellular responses such as fate decision, cell survival/death, secretion, and migration. For example, morphogens are secreted signaling molecules that form concentration gradients within tissues and induce distinct cell fates in a signal strength-dependent manner. In addition to extracellular ligand abundance, the sensitivity of signal-receiving cells to ligands also influences signal strength. Cell sensitivity to ligands is controlled at various levels: receptor presentation at the cell surface, positive/negative regulation of signal transduction, and target gene activation/repression. While the regulation of signal transduction and gene transcription is well studied, receptor presentation is still not fully understood. Recently, it was reported that cellular sensitivity to the Wingless (Wg)/Wnt morphogen is regulated by balanced ubiquitination and deubiquitination of its receptor Frizzled (Fz). In this review, we review how ubiquitination regulates receptor presentation at the cell surface for the detection of extracellular signal strength.

      DOI: 10.1007/s00018-012-1084-4

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    • 糖鎖関連遺伝子の大規模スクリーニング

      山本(日野)美紀, 山本(日野)美紀, 蟹江善美, 粟野若枝, 木下聖子, 西原祥子, 岡野栄之, 上田龍, 蟹江治, 蟹江治, 後藤聡, 後藤聡

      生化学   2011年

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    • 受容体のユビキチン化と脱ユビキチン化を介した細胞のWnt応答性の制御 査読有り

      駒田雅之, 後藤聡

      細胞工学29   1237 - 1243   2010年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    • 糖鎖修飾の場—ゴルジ体ユニットによる糖鎖修飾の制御 招待有り 査読有り

      後藤聡

      蛋白質核酸酵素53   1475 - 1479   2008年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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    • ショウジョウバエにおけるメンブレントラフィック 招待有り 査読有り

      阿部将人, 後藤聡

      蛋白質核酸酵素53   2193 - 2199   2008年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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    • ショウジョウバエを用いた糖鎖修飾制御因子のゲノムワイドスクリーニング

      山本(日野)美紀, 矢野弘之, 蟹江善美, 粟野若枝, 平井ゆう, 桑原玲子, 木下(青木)聖子, 上田龍, 西原祥子, 西原祥子, 蟹江治, 後藤聡, 後藤聡

      生化学   2007年

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    • Regulation of glycosylation by golgi units 招待有り 査読有り

      Satoshi Goto

      Trends in Glycoscience and Glycotechnology18 ( 104 ) 347 - 351   2006年

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      記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:Gakushin Publishing Company  

      Most secretory and transmembrane proteins are post-translationally modified with various glycans. The structures of glycans attached to the proteins depend on proteins, cells, tissues, ages, species and individuals. However, the mechanism of regulation to generate such diverse glycans in a cell has not been revealed. In this review, we would like to discuss our finding, "Golgi units" that would regulate the variety of glycosylation. ©2006 FCCA (Forum: Carbohydrates Coming of Age).

      DOI: 10.4052/tigg.18.347

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    • RNAi変異体ハエバンクを用いたショウジョウバエ遺伝子機能解析

      藤谷和子, 佐渡由希子, 高橋邦明, 内藤雄樹, 山田智之, 森下真一, 西郷薫, 後藤聡, 西原祥子, 上田龍

      日本分子生物学会年会講演要旨集28th   543   2005年11月25日

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      記述言語:日本語  

      J-GLOBAL

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    • Functional glycomics using Drosophila RNAi system

      S Nishihara, M Hino, H Yoshida, N Sasaki, S Goto, H Toyoda, R Ueda

      GLYCOBIOLOGY14 ( 11 ) 1065 - 1065   2004年11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:OXFORD UNIV PRESS INC  

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    • Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter

      S Kamiyama, T Suda, R Ueda, H Yoshida, N Kikuchi, Y Chiba, S Goto, H Toyoda, H Narimatsu, Y Jigami, S Nishihara

      GLYCOBIOLOGY13 ( 11 ) 857 - 857   2003年11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:OXFORD UNIV PRESS INC  

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    • PAPSトランスポーターのクローニングと機能解析~PAPSトランスポーターは生命維持に必須である

      神山伸, 須田健, 上田龍, 浜本洋, 鈴木正之, 菊池紀広, 千葉靖典, 後藤聡, 西原祥子

      日本糖質学会年会要旨集24th   52   2003年7月3日

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      記述言語:日本語  

      J-GLOBAL

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    • 発生のおける小胞輸送の役割 招待有り 査読有り

      後藤聡

      実験医学21   136 - 141   2003年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    • ショウジョウバエ脚のパターン形成、近遠軸形成機構 招待有り 査読有り

      後藤聡, 林茂生

      実験医学17   306 - 312   1999年

      詳細を見る

      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    • 発生研究のための蛍光プローブによるmRNAとタンパク質の二重染色法 招待有り 査読有り

      後藤聡, 林茂生

      実験医学16   523 - 527   1998年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    • Dpp, Wgによるショウジョウバエの翅・肢誘導と近遠軸形成 招待有り 査読有り

      後藤聡, 林茂生

      実験医学15   342 - 349   1997年

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      記述言語:日本語   掲載種別:記事・総説・解説・論説等(その他)  

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    書籍等出版物

    • Localization of glycosyl enzymes and nucleotide-sugar transporters in the endoplasmic reticulum and the Golgi apparatus

      Yamamoto-Hino,M, Goto,S( 担当: 共著)

      SpringerReference, Heidelberg  2014年 

      詳細を見る

      記述言語:英語 著書種別:その他

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    • 「ゴルジ体(糖修飾の場として)」 生化学辞典

      後藤聡( 担当: 共著)

      朝倉書店  2012年 

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      記述言語:日本語 著書種別:事典・辞書

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    • 「EGF経路」「ERK/MAPK経路」「Hedgehogシグナル」「AKT」 岩波生物学辞典

      後藤聡( 担当: 共著)

      岩波書店  2012年 

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      記述言語:日本語 著書種別:事典・辞書

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    • Structure, Function and Formation of Glycans in Drosophila.

      Yamamoto-Hino,M, Okano,H, Kanie,O, Goto,S( 担当: 共著)

      Nova Science publishers, Inc., NY  2012年 

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      記述言語:英語 著書種別:学術書

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    • 5. 「成虫原基」新生化学実験講座「発生・分化・老化」

      後藤聡, 谷村禎一( 担当: 共著)

      東京化学同人  1992年 

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      記述言語:日本語 著書種別:事典・辞書

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    共同研究・競争的資金等の研究

    • 糖鎖およびリン酸修飾の基盤となる選別輸送ゾーンの分子機構と生理機能の解析

      文部科学省  科学研究費助成事業 

      詳細を見る

      2017年7月 - 2022年3月

      資金種別:競争的資金

      配分額:17900000円 ( 直接経費:12530000円 、 間接経費:5370000円 )

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    • 核膜と小胞体の機能的連携

      日本学術振興会  科学研究費助成事業 

      詳細を見る

      2016年4月 - 2017年3月

      資金種別:競争的資金

      配分額:2800000円 ( 直接経費:1960000円 、 間接経費:840000円 )

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    • GPI修飾の新規メカニズムとその破綻によるシグナル伝達異常の解明

      文部科学省  科学研究費助成事業 

      詳細を見る

      2013年4月 - 2015年3月

      資金種別:競争的資金

      配分額:8200000円 ( 直接経費:5740000円 、 間接経費:2460000円 )

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    • 自然免疫をダイナミックに制御する糖鎖修飾の解析

      日本学術振興会  科学研究費助成事業 

      詳細を見る

      2012年4月 - 2014年4月

      資金種別:競争的資金

      配分額:14100000円 ( 直接経費:9870000円 、 間接経費:4230000円 )

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    • mRNAの局在による翻訳後修飾の制御

      文部科学省  科学研究費助成事業 

      詳細を見る

      2011年4月 - 2013年3月

      資金種別:競争的資金

      配分額:7200000円 ( 直接経費:5040000円 、 間接経費:2160000円 )

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    • 糖鎖修飾による自然免疫システムの多様な制御機構

      住友財団  住友財団、基礎科学研究助成 

      詳細を見る

      2011年4月 - 2013年3月

      資金種別:競争的資金

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