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 】

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    The RNA Society 

  • 　
    　
    　
     

    Genetics Society of America 

  • 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 】

  • Profiling of RBM20-Regulated CaMKIIδ Splice Variants Across the Heart, Skeletal Muscle, and Olfactory Bulbs.

    Yui MAEDA, Yuri YAMASU and Hidehito KUROYANAGI

    Genes to Cells ( John Wiley & Sons, Inc. )    2025 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

    　View Summary

    Calcium/calmodulin-dependent protein kinase IIδ (CaMKIIδ), encoded by the Camk2d gene, plays key regulatory roles in various Ca2+-regulated cellular processes. Extensive alternative splicing of the Camk2d gene generates multiple CaMKIIδ splice variants that exhibit differential roles. Despite significant advances in understanding the functions of CaMKIIδ, the full repertoire of Camk2d splice variants in a variety of tissues and their distinct roles in physiological and pathological contexts remain incompletely characterized due to the complex nature of multiple alternative splicing events. Here, we conducted long-read amplicon sequencing to investigate the murine Camk2d splice variants in the heart, skeletal muscle, and olfactory bulbs and show that Camk2d mRNAs in the heart and skeletal muscle have shorter 3'UTRs. Our results in this study suggest that a key regulator of Camk2d splicing, RNA-binding motif protein 20 (RBM20), whose gain-of-function mutations cause dilated cardiomyopathy, is crucial for the expression of heart-specific splice variants. Olfactory bulbs specifically express novel splice variants that utilize a mutually exclusive exon 6B and/or an alternative polyadenylation site in a novel exon 17.5 in an RBM20-inependent manner. The tissue-specific repertoire of CaMKIIδ splice variants and their aberrant expression in disease model animals will help in understanding their roles in physiological and pathological contexts.

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      DOI

  • 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.

  • 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.

  • Comprehensive analysis of alternative splicing coupled with nonsense-mediated mRNA decay (AS-NMD) in C. elegans

    Kuroyanagi Hidehito

      94 ( 6 ) 868 - 874   2022.12 [ Peer Review Accepted ]

    Type of publication: Research paper (scientific journal)

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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 】

  • Elucidation of pathogenesis mechanisms and exploration of therapeutic strategies using mouse models of dilated cardiomyopathy

    Challenging research (sprout)

    Project Year: 2023.06  -  2025.03 

    Investigator(s): KUROYANAGI Hidehito 

    Direct: 5,000,000 (YEN)  Overheads: 1,500,000 (YEN)  Total: 6,500,000 (YEN)

  • Elucidation of pathogenesis mechanisms and exploration of therapeutic strategies using mouse models of dilated cardiomyopathy

    Challenging research (sprout)

    Project Year: 2023.06  -  2025.03 

    Direct: 5,000,000 (YEN)  Overheads: 6,500,000 (YEN)  Total: 1,500,000 (YEN)

  • Functional Analysis of RBM20 Whose Mutation Causes Dilated Cardiomyopathy

    Challenging research (sprout)

    Project Year: 2020.07  -  2023.03 

    Investigator(s): KUROYANAGI Hidehito 

    Direct: 5,000,000 (YEN)  Overheads: 1,500,000 (YEN)  Total: 6,500,000 (YEN)

  • Tissue-specific pre-mRNA processing in living animals

    Grant-in-Aid for Scientific Research(B)

    Project Year: 2020.04  -  2023.03 

    Investigator(s): KUROYANAGI Hidehito 

    Direct: 13,700,000 (YEN)  Overheads: 4,110,000 (YEN)  Total: 17,810,000 (YEN)

  • Tissue-specific pre-mRNA processing in living animals

    Grant-in-Aid for Scientific Research(B)

    Project Year: 2020.04  -  2023.03 

    Direct: 13,700,000 (YEN)  Overheads: 17,810,000 (YEN)  Total: 4,110,000 (YEN)

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Social Activity 【 display / non-display 】

  • Special Lecture & Lab Exercise in Okinawa AMICUS International Junior High School

    Okinawa AMICUS International Junior High School 

    2024.09
    　
    　

  • Ninufa-Bushi Seminar

    University of the Ryukyus  The 5th Ninufa-Bushi Seminar in the University of the Ryukyus 

    2023.08
    　
    　

  • Ninufa-Bushi Seminar

    University of the Ryukyus  The 4th Ninufa-Bushi Seminar in the University of the Ryukyus 

    2022.08
    　
    　

  • NHK TV program presentes our research findings

    NHK  NHK BSP/BS4K TV program "HUMANIENCE" 

    2022.07
    -
    2022.08

  • University of the Ryukyus 

    2021.11
    　
    　

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Media Coverage 【 display / non-display 】

  • 細胞や遺伝子の仕組み学ぶ　うるま・アミークス　血液型判定実験も  Newspaper, magazine

    琉球新報  2024.10

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    沖縄県うるま市にある沖縄アミークスインターナショナル中学校で黒柳秀人教授が行った特別講義と分子生物学実習の様子が、琉球新報で紹介されました。

  • 琉球大学で高校生が医学部の体験授業  TV or radio program

    NHK沖縄放送局  NHKニュース  2023.8

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    2023年8月21日（月）～22日（火）に琉球大学医学部で行われた第5回「琉大にぬふぁ星講座（医学部体験授業）」が、NHKテレビのニュースで紹介されました。生化学講座は、PCR法と制限酵素消化により血液型判定を行う実験実習①の担当講座のひとつとして、スタッフ全員で参加しました。

  • 高校生、医学部授業を体験　琉大が講座　本物の機器使い実習も  Newspaper, magazine

    沖縄タイムス  2022.8

    Author： Other 

  • 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。

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