Current Affiliation Organization 【 display / non-display 】

Current Affiliation Organization 【 display / non-display 】

University 【 display / non-display 】

University 【 display / non-display 】

Graduate School 【 display / non-display 】

Graduate School 【 display / non-display 】

Academic degree 【 display / non-display 】

Academic degree 【 display / non-display 】

External Career 【 display / non-display 】

External Career 【 display / non-display 】

display all >>

Affiliated academic organizations 【 display / non-display 】

Affiliated academic organizations 【 display / non-display 】

Research Interests 【 display / non-display 】

Research Interests 【 display / non-display 】

display all >>

Research Areas 【 display / non-display 】

Research Areas 【 display / non-display 】

Published Papers 【 display / non-display 】

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)

  　View Summary

  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.

  • Access this article

    DOI

• 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)

  　View Summary

  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.

  • Access this article

    DOI

• 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)

  　View Summary

  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.

  • Access this article

    DOI

• 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)

  　View Summary

  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.

  • Access this article

    DOI

• 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)

  　View Summary

  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.

  • Access this article

    DOI

display all >>

Books 【 display / non-display 】

Books 【 display / non-display 】

• Encyclopedia of Genetics

  ( Part: Multiple Authorship )

  2022.01

• Alternative pre-mRNA Splicing: Theory and Protocols

  ( Part: Allotment Writing )

  Wiley-VCH  2012.01

Other Papers 【 display / non-display 】

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

Presentations 【 display / non-display 】

Presentations 【 display / non-display 】

• 拡張型心筋症の原因となるスプライシング制御因子RBM20の変異解析とモデルマウスの作製

  黒柳秀人, 井原健介, 大野, 都甲麻理奈, 平岡優一, 木村彰方, 笹野哲郎

  第23回日本心不全学会学術集会  (広島市)  2019.10  -  2019.10 

• Alternative splicing regulation of the S-adenosylmethionine synthase (sams) genes through adenosine methylation at their 3’ splice sites

  Shotaro Wani, Eichi Watabe, Hidehito Kuroyanagi

  線虫研究の未来を創る会 2019  (名古屋大学)  2019.08  -  2019.08 

• Nascent RNA sequencing reveals pre-mRNA processing dynamics governed by splicing regulators

  Eichi Watabe, Hidehito Kuroyanagi

  線虫研究の未来を創る会 2019  (名古屋大学)  2019.08  -  2019.08 

• PES-4 regulates head-muscle-specific alternative splicing of the tropomyosin pre-mRNA

  Eichi Watabe, Hidehito Kuroyanagi

  The 22nd International C. elegans Meeting  (Los Angeles, CA)  2019.06  -  2019.06 

• Genome-Wide Kinetic Analysis of pre-mRNA Processing in C. elegans

  Eichi Watabe, Hidehito Kuroyanagi

  The 22nd International C. elegans Meeting  (Los Angeles, CA)  2019.06  -  2019.06 

display all >>

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

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)

• 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)

• 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)

display all >>

Social Activity 【 display / non-display 】

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

  　

  　

  

display all >>

Media Coverage 【 display / non-display 】

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

  　View Summary

  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

  　View Summary

  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 

  　View Summary

  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 

  　View Summary

  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。

display all >>

Academic Activities 【 display / non-display 】

Academic Activities 【 display / non-display 】

• [International contribution]

  KUROYANAGI Hidehito  （ NAHA Cultural Arts Theater, NAHArt ）

  2023.7

  　

  　

  種別： Competition, symposium, etc.