KUROYANAGI Hidehito

写真a

Title

Professor

Researcher Number(JSPS Kakenhi)

30323702

Date of Birth

1970

Laboratory Address

207 Uehara, Nishihara-cho, Okinawa, Japan

Mail Address

E-mail address

Laboratory Phone number

+81-98-895-1112

Homepage URL

https://biochem.med.u-ryukyu.ac.jp/en/

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Current Affiliation Organization 【 display / non-display 】

  • Duty   University of the Ryukyus   Graduate School of Medicine   Professor  

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University 【 display / non-display 】

  • 1989.04
    -
    1994.03

    The University of Tokyo   Faculty of Science   Graduated

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Graduate School 【 display / non-display 】

  • 1994.04
    -
    1996.03

    The University of Tokyo  Graduate School, Division of Science  Master's Course  Completed

  • 1996.04
    -
    1999.03

    The University of Tokyo  Graduate School, Division of Science  Doctor's Course  Completed

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Academic degree 【 display / non-display 】

  • The University of Tokyo -  PhD in Science

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External Career 【 display / non-display 】

  • 1997.04
    -
    1999.03

     

  • 1999.04
    -
    2000.03

     

  • 2000.04
    -
    2003.08

    Tokyo Medical and Dental University, Assistant Professor  

  • 2003.09
    -
    2008.03

    Tokyo Medical and Dental University, Junior Associate Professor  

  • 2008.04
    -
    2012.03

    Tokyo Medical and Dental University, Associate Professor  

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Affiliated academic organizations 【 display / non-display 】

  • 　
    　
    　
     

    Genetics Society of America 

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    The RNA Society 

  • 2016.04
    -
    2018.03
     

    The RNA Society of Japan    Council Officer

  • 2018.04
    -
    2020.03
     

    The RNA Society of Japan    Council Officer

  • 2020.04
    -
    2022.03
     

    The RNA Society of Japan    Council Officer

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Research Interests 【 display / non-display 】

  • Molecular Biology

  • transcriptome

  • model organism

  • gene expression

  • development

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Research Areas 【 display / non-display 】

  • Life Science / Molecular biology

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Published Papers 【 display / non-display 】

  • Alternative splicing of a single exon causes a major impact on the affinity of Caenorhabditis elegans tropomyosin isoforms for actin filaments.

    Shoichiro Ono, Eichi Watabe, Keita Morisaki, Kanako Ono, Hidehito Kuroyanagi

    Frontiers in Cell and Developmental Biology ( Frontiers Media S.A. )  11   1208913   2023.09 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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    Tropomyosin is generally known as an actin-binding protein that regulates actomyosin interaction and actin filament stability. In metazoans, multiple tropomyosin isoforms are expressed, and some of them are involved in generating subpopulations of actin cytoskeleton in an isoform-specific manner. However, functions of many tropomyosin isoforms remain unknown. Here, we report identification of a novel alternative exon in the #ICaenorhabditis elegans#IR tropomyosin gene and characterization of the effects of alternative splicing on the properties of tropomyosin isoforms. Previous studies have reported six tropomyosin isoforms encoded by the #IC. elegans lev-11#IR tropomyosin gene. We identified a seventh isoform, LEV-11U, that contained a novel alternative exon, exon 7c (E7c). LEV-11U is a low-molecular-weight tropomyosin isoform that differs from LEV-11T only at the exon 7-encoded region. #IIn silico#IR analyses indicated that the E7c-encoded peptide sequence was unfavorable for coiled-coil formation and distinct from other tropomyosin isoforms in the pattern of electrostatic surface potentials. #IIn vitro#IR, LEV-11U bound poorly to actin filaments, whereas LEV-11T bound to actin filaments in a saturable manner. When these isoforms were transgenically expressed in the #IC. elegans#IR striated muscle, LEV-11U was present in the diffuse cytoplasm with tendency to form aggregates, whereas LEV-11T co-localized with sarcomeric actin filaments. Worms with a mutation in E7c showed reduced motility and brood size, suggesting that this exon is important for the optimal health. These results indicate that alternative splicing of a single exon can produce biochemically diverged tropomyosin isoforms and suggest that a tropomyosin isoform with poor actin affinity has a novel biological function.

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  • Simultaneous measurement of nascent transcriptome and translatome using 4-thiouridine metabolic RNA labeling and translating ribosome affinity purification.

    Hirotatsu Imai, Daisuke Utsumi, Hidetsugu Torihara, Kenzo Takahashi, Hidehito Kuroyanagi, Akio Yamashita

    Nucleic Acids Research ( Oxford University Press )  51 ( 14 ) e76   2023.06 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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    Regulation of gene expression in response to various biological processes, including extracellular stimulation and environmental adaptation requires nascent RNA synthesis and translation. Analysis of the coordinated regulation of dynamic RNA synthesis and translation is required to determine functional protein production. However, reliable methods for the simultaneous measurement of nascent RNA synthesis and translation at the gene level are limited. Here, we developed a novel method for the simultaneous assessment of nascent RNA synthesis and translation by combining 4-thiouridine (4sU) metabolic RNA labeling and translating ribosome affinity purification (TRAP) using a monoclonal antibody against evolutionarily conserved ribosomal P-stalk proteins. The P-stalk-mediated TRAP (P-TRAP) technique recovered endogenous translating ribosomes, allowing easy translatome analysis of various eukaryotes. We validated this method in mammalian cells by demonstrating that acute unfolded protein response (UPR) in the endoplasmic reticulum (ER) induces dynamic reprogramming of nascent RNA synthesis and translation. Our nascent P-TRAP (nP-TRAP) method may serve as a simple and powerful tool for analyzing the coordinated regulation of transcription and translation of individual genes in various eukaryotes.

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  • Structure of the Caenorhabditis elegans m6A methyltransferase METT10 that regulates SAM homeostasis.

    Jue Ju, Tomohiko Aoyama, Yuka Yashiro, Seisuke Yamashita, Hidehito Kuroyanagi, Kozo Tomita

    Nucleic Acids Research ( Oxford University Press )  51 ( 5 ) 2434 - 2446   2023.02 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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    In #ICaenorhabditis elegans#IR, the N#U6#UR-methyladenosine (m#U6#URA) modification by METT10, at the 3'-splice sites in #IS#IR-adenosyl-L-methionine (SAM) synthetase (#Isams#IR) precursor mRNA (pre-mRNA), inhibits #Isams#IR pre-mRNA splicing, promotes alternative splicing coupled with nonsense-mediated decay of the pre-mRNAs, and thereby maintains the cellular SAM level. Here, we present structural and functional analyses of #IC. elegans#IR METT10. The structure of the N-terminal methyltransferase domain of METT10 is homologous to that of human METTL16, which installs the m#U6#URA modification in the 3'-UTR hairpins of methionine adenosyltransferase (#IMAT2A#IR) pre-mRNA and regulates the MAT2A pre-mRNA splicing/stability and SAM homeostasis. Our biochemical analysis suggested that #IC. elegans#IR METT10 recognizes the specific structural features of RNA surrounding the 3'-splice sites of #Isams#IR pre-mRNAs, and shares a similar substrate RNA recognition mechanism with human METTL16. #IC. elegans#IR METT10 also possesses a previously unrecognized functional C-terminal RNA-binding domain, kinase associated 1 (KA-1), which corresponds to the vertebrate-conserved region (VCR) of human METTL16. As in human METTL16, the KA-1 domain of #IC. elegans#IR METT10 facilitates the m#U6#URA modification of the 3'-splice sites of #Isams#IR pre-mRNAs. These results suggest the well-conserved mechanisms for the m#U6#URA modification of substrate RNAs between #IHomo sapiens#IR and #IC. elegans#IR, despite their different regulation mechanisms for SAM homeostasis.

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  • I536T variant of RBM20 affects splicing of cardiac structural proteins that are causative for developing dilated cardiomyopathy.

    Takuma Yamamoto, Rie Sano, Aya Miura, Mai Imasaka, Yoshiro Naito, Minori Nishiguchi, Kensuke Ihara, Naruhito Otani, Yoshihiko Kominato, Masaki Ohmuraya, Hidehito Kuroyanagi & Hajime Nishio

    Journal of Molecular Medicine ( Springer Nature )    2022.10 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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    RBM20 is one of the genes predisposing to dilated cardiomyopathy (DCM). Variants in the RS domain have been reported in many DCM patients, but the pathogenicity of variants within the RNA-recognition motif remains unknown. Two human patients with the I536T-RBM20 variant without an apparent DCM phenotype were identified in sudden death cohorts. A splicing reporter assay was performed, and an I538T knock-in mouse model (Rbm20I538T) was generated to determine the significance of this variant. The reporter assay demonstrated that the human I536T variant affected the TTN splicing pattern compared to wild-type. In the mouse experiments, Rbm20I538T mice showed different splicing patterns in Ttn, Ldb3, Camk2d, and Ryr2. The expressions of Casq1, Mybpc2, and Myot were upregulated in Rbm20I538T mice, but Rbm20I538T mice showed neither DCM nor cardiac dysfunction on histopathological examination and ultrasound echocardiography. The I536T-RBM20 (I538T-Rbm20) variant changes gene splicing and affects gene expression, but the splicing and expression changes in Ttn and Ca handling genes such as Casq1, Camk2d, and Ryr2 do not cause DCM morphology in the mouse model.

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  • Periostin Exon-21 Antibody Neutralization of Triple-Negative Breast Cancer Cell-Derived Periostin Regulates Tumor-Associated Macrophage Polarization and Angiogenesis.

    Tatsuya Fujikawa, Fumihiro Sanada, Yoshiaki Taniyama, Kana Shibata, Naruto Katsuragi, Nobutaka Koibuchi, Kaori Akazawa, Yuko Kanemoto, Hidehito Kuroyanagi, Kenzo Shimazu, Hiromi Rakugi and Ryuichi Morishita

    Cancers   13 ( 20 ) 5072   2021.10 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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    Periostin (Pn) is involved in multiple processes of cancer progression. Previously, we reported that Pn expression is correlated with mesenchymal tumor markers and poor prognosis in triple-negative breast cancer (TNBC). In the TNBC xenograft model, chemotherapy increased expression of a Pn alternative splicing variant (ASV) with exon 21, and administration of the neutralizing antibody against Pn with exon 21 (Pn-21 Ab) overcame chemoresistance with a reduction in the mesenchymal cancer cell fraction. In the present study, the role of Pn ASV with exon 21 in TNBC progression has been addressed. We first established a stable cell line carrying a fluorescence-based splicing reporter. Pn-positive TNBC has higher expression of genes related to tumor-associated macrophage (TAM) recruitment and ECM-receptor interaction than Pn-negative cells. In a xenograft model, only Pn-positive cells initiated tumor formation, and the Pn-21 Ab suppressed tumor cell growth, accompanied by decreased M2 TAM polarization and the number of tumor vessels. These data suggest that cancer cell-derived Pn ASV educates TAMs and regulates angiogenesis, which in turn establishes a microenvironmental niche that is supportive of TNBC.

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Books 【 display / non-display 】

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Other Papers 【 display / non-display 】

  • New approaches to decipher pre-mRNA splicing codes

    Hidehito Kuroyanagi

    Seikagaku   82 ( 5 ) 402 - 411   2010  [Refereed]

     

    PubMed

  • Visualization of alternative splicing events in vivo.

    Hidehito Kuroyanagi

    Tanpakushitu Kakusan Koso   54 ( 16 ) 2044 - 2048   2009.12  [Refereed]

     

    PubMed

  • Structure, expression and functions of receptor-type protein tyrosine phosphatase RPTP-BK in central nervous system

    Hidehito Kuroyanagi, Masatoshi Tagawa, Takuji Shirasawa

    Tanpakushitu Kakusan Koso   43 ( 8 ) 1162 - 1168   1998.06  [Refereed]

     

    PubMed

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Presentations 【 display / non-display 】

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Grant-in-Aid for Scientific Research 【 display / non-display 】

  • Grant-in-Aid for Scientific Research(B)

    Project Year: 2017  -  2019 

    Direct: 0 (YEN)  Overheads: 0 (YEN)  Total: 0 (YEN)

  • Grant-in-Aid for Scientific Research(B)

    Project Year: 2014  -  2017 

    Direct: 0 (YEN)  Overheads: 0 (YEN)  Total: 0 (YEN)

  • Grant-in-Aid for Scientific Research(B)

    Project Year: 2014  -  2016 

    Direct: 0 (YEN)  Overheads: 0 (YEN)  Total: 0 (YEN)

  • Grant-in-Aid for challenging Exploratory Research

    Project Year: 2014  -  2015 

    Direct: 0 (YEN)  Overheads: 0 (YEN)  Total: 0 (YEN)

  • Grant-in-Aid for challenging Exploratory Research

    Project Year: 2014  -  2015 

    Direct: 0 (YEN)  Overheads: 0 (YEN)  Total: 0 (YEN)

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Social Activity 【 display / non-display 】

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Media Coverage 【 display / non-display 】

  • 高校生ら医学部体験　琉大　進学支援で模擬授業  Newspaper, magazine

    沖縄タイムス社  沖縄タイムス  2021.11

    Author： Other 

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    医師や医学系研究者を志す高校生に医学部で学ぶ内容を知ってもらおうと、琉球大学は１０月２３、２４の両日、同大で模擬体験授業を行った。高校生１５人が参加し、実際に授業で使われる医療機器などを用いて講義を受けた。

  • 八重高生２人「意欲高まった」　琉大医学部で体験授業  Newspaper, magazine

    八重山日報社  八重山日報  教育, 教育・スポーツ  2021.10

    Author： Other 

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    琉球大学医学部は２３、２４の両日、同学部の体験授業「琉球大学にぬふぁ星講座」を県内の高校生向けに開講した。

  • 心筋症の原因遺伝子をｉＰＳで確認…阪大など  Newspaper, magazine

    読売新聞大阪本社  読売新聞  関西発  2021.5

    Author： Other 

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    大阪大学の研究チームが英科学誌に発表した研究成果に対するコメント

  • RNAの加工パターンを動物が生きたまま観察する手法を開発。遺伝子発現のしくみに迫る 黒柳秀人先生 東京医科歯科大学  Internet

    学校法人河合塾  みらいぶプラス  ＜専門分野：分子生物学＞  2017.4

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    遺伝子がはたらくしくみは、ヒトを含む哺乳類と線虫やショウジョウバエなどのモデル動物でとてもよく似ている。黒柳先生は、蛍光タンパク質を使ってミニ遺伝子を作製、モデル生物に導入し、RNAの加工パターンを生きたまま観察する方法の開発に成功した。一つの遺伝子から多様なRNAやタンパク質が作られるしくみを解明し、複雑なヒトの遺伝子発現制御のしくみやその破綻による疾患の解明につなげたいと考える。

  • タンパク質が2つ協働してRNA認識  Internet

    科学技術振興機構（JST）  サイエンスポータル  ニュース - 速報・レビュー（ニュース速報） -  2014.8

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    2種類のタンパク質がメッセンジャー(m)RNA前駆体を協働的に認識して選択的プロセシング(加工)を制御し、筋肉で働くように加工していることを、東京医科歯科大学難治疾患研究所の黒柳秀人(ひでひと)准教授らが線虫で初めて確かめた。同様のRNAの認識、制御、加工の仕組みは哺乳類を含む生物に広く存在するとみられている。

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