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Identification of the minimalized essential unit for genetic inheritance of artificial mini-chromosomes

Research Project

Project/Area Number 19K22402
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionKindai University

Principal Investigator

Shinohara Miki  近畿大学, 農学部, 教授 (80335687)

Project Period (FY) 2019-06-28 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2021: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords減数分裂期 / 相同組換え / 人工染色体 / 染色体分配 / 組換え
Outline of Research at the Start

減数分裂期は子孫へ遺伝情報を継承する唯一の機構である。減数分裂期交叉型組換えは、染色体の正確な分配と遺伝情報多様性の創出に機能する。その反応は組換え酵素、染色体高次構造も関わり、厳密な制御の下に行われることから、この制御機構を明らかにする。出芽酵母人工ミニ染色体を用いて、減数分裂期染色体分配に特化した新たな必須最小ユニットについて、DNA配列や染色体機能の必要条件を合成生物学的手法を用いて解析を行う。染色体レベルでの減数分裂期過程の理解が進めば、その延長上にある細胞老化のリセットの機構、遺伝的多様性創出、そして異種と同種を分ける機構の理解など、減数分裂期が持つ重要な機能の理解につながる。

Outline of Final Research Achievements

In eukaryotes, Saccharomyces cerevisiae allows the construction of fully artificially synthesized chromosomes using the minimum regions of the basic units of chromosomal function such as replication origin, centromeres, and telomeres. Here we show that the artificial mini-chromosomes show frequent non-disjunction of homologs during the first division of meiosis while normal segregation during the second division. In addition, since the cause of non-disjunction was considered to be meiotic crossover recombination deficiency, UAS sequence of the GAL1 promoter was inserted on the artificial mini-chromosome and analyzed chromosome segregation in Gal4-BD-Spo11 expressing strains, but no improvement was observed. On the other hand, autophosphorylation-mediated activation of the chromosomal axis factor Mek1 kinase plays an important role in template selection for meiotic recombination, revealing that ectopic recombination occurs frequently in the phosphorylation-deficient mek1 mutant.

Academic Significance and Societal Importance of the Research Achievements

1真核生物の中でセントロメア、テロメア、複製開始点など染色体の必須要素が明らかになっている出芽酵母で、人工染色体にさまざまな人工DNA配列を付加することによって引き起こされる減数分裂期組換えの挙動の変化について解析を行うことで、細胞内における染色体サイズの認識とそれに応じた交叉型組換え制御機構について明らかにし、次世代に継承可能にするための最小のゲノム要素を明らかにすることができる。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (34 results)

All 2022 2021 2020 2019 Other

All Int'l Joint Research (8 results) Journal Article (7 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 7 results,  Open Access: 7 results) Presentation (17 results) (of which Int'l Joint Research: 1 results,  Invited: 6 results) Remarks (2 results)

  • [Int'l Joint Research] Chung-Ang University(韓国)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] Indian Inst. of Sci Education and Res.(インド)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] European Molecular Biology Laboratory(ドイツ)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] Stanford University(米国)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] Chung-Ang University(韓国)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] Rutgers New Jersey Medical School(米国)

    • Related Report
      2020 Research-status Report
  • [Int'l Joint Research] University of Chicago/Rutgers New Jersey Medical School(米国)

    • Related Report
      2019 Research-status Report
  • [Int'l Joint Research] FMI(スイス)

    • Related Report
      2019 Research-status Report
  • [Journal Article] The synaptonemal complex central region modulates crossover pathways and feedback control of meiotic double-strand break formation2021

    • Author(s)
      Lee Min-Su、Higashide Mika T、Choi Hyungseok、Li Ke、Hong Soogil、Lee Kangseok、Shinohara Akira、Shinohara Miki、Kim Keun?P
    • Journal Title

      Nucleic Acids Research

      Volume: 49 Issue: 13 Pages: 7537-7553

    • DOI

      10.1093/nar/gkab566

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Regulation of Msh4-Msh5 association with meiotic chromosomes in budding yeast2021

    • Author(s)
      Nandanan Krishnaprasad G、Salim Sagar、Pankajam Ajith V、Shinohara Miki、Lin Gen、Chakraborty Parijat、Farnaz Amamah、Steinmetz Lars M、Shinohara Akira、Nishant Koodali T
    • Journal Title

      Genetics

      Volume: 219 Issue: 2

    • DOI

      10.1093/genetics/iyab102

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Phosphorylation of luminal region of the SUN-domain protein Mps3 promotes nuclear envelope localization during meiosis2021

    • Author(s)
      Prasada Rao Hanumanthu BD、Sato Takeshi、Challa Kiran、Fujita Yurika、Shinohara Miki、Shinohara Akira
    • Journal Title

      eLife

      Volume: 10 Pages: 1-29

    • DOI

      10.7554/elife.63119

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] SCFCdc4 ubiquitin ligase regulates synaptonemal complex formation during meiosis.2020

    • Author(s)
      Zhihui Zhu, Mohammad Bani Ismail, Miki Shinohara, Akira Shinohara
    • Journal Title

      Life science alliance

      Volume: 4(2) Issue: 2 Pages: e202000933-e202000933

    • DOI

      10.26508/lsa.202000933

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genetic interactions of histone modification machinery, Set1 and PAF1C, with the recombination complex, Rec114-Mer2-Mei4, in the formation of meiotic DNA double-strand breaks2020

    • Author(s)
      69.Zhang Y., Suzuki T., Li K., Gothwal S.K., Shinohara M., and A. Shinohara
    • Journal Title

      International Journal of Molecular Sciences

      Volume: in press Issue: 8 Pages: 2679-2679

    • DOI

      10.3390/ijms21082679

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Enhanced homologous recombination by the modulation of targeting vector ends2020

    • Author(s)
      Hirotsune Shinji、Kiyonari Hiroshi、Jin Mingyue、Kumamoto Kanako、Yoshida Kayo、Shinohara Miki、Watanabe Hitomi、Wynshaw-Boris Anthony、Matsuzaki Fumio
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 1-12

    • DOI

      10.1038/s41598-020-58893-9

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Distinct Functions in Regulation of Meiotic Crossovers for DNA Damage Response Clamp Loader Rad24(Rad17) and Mec1(ATR) Kinase2019

    • Author(s)
      Shinohara Miki、Bishop Douglas K.、Shinohara Akira
    • Journal Title

      Genetics

      Volume: 213 Issue: 4 Pages: 1255-1269

    • DOI

      10.1534/genetics.119.302427

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 減数分裂期特異的な染色体軸-ループ構造による減数分裂期 組換え制御2022

    • Author(s)
      篠原美紀
    • Organizer
      蛋白研セミナー:生殖細胞・減数分裂研究の過去・現在・未来/生殖細胞・減数分裂研究の最前線
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Sae2のDNA ligase VIを介したNHEJ抑制機構の分子メカニズム2021

    • Author(s)
      松嵜健一郎, 森田一世, 篠原美紀
    • Organizer
      日本遺伝学会,第93回大会_東京2021
    • Related Report
      2021 Annual Research Report
  • [Presentation] Analysis of the regulatory mechanism through Rad50 for bi-directional DSB resection.2021

    • Author(s)
      玉井智貴, 森田一世, 篠原美紀
    • Organizer
      日本遺伝学会,第93回大会_東京2021
    • Related Report
      2021 Annual Research Report
  • [Presentation] 出芽酵母における DNA二重鎖切断修復経路での Pso2ヌクレアーゼの機能の解析2021

    • Author(s)
      笠井公輔、 篠原美紀
    • Organizer
      日本遺伝学会,第93回大会_東京2021
    • Related Report
      2021 Annual Research Report
  • [Presentation] Budding Yeast Protein Phosphatase 4 Promoted Meiotic Axes Formation through Hop1 Assembly2021

    • Author(s)
      Ke Li, Miki Shinohara
    • Organizer
      第26回DNA複製・組換え・修復ワークショップ
    • Related Report
      2021 Annual Research Report
  • [Presentation] 両方向的DSB末端単鎖化のRad50による制御機構の解析2021

    • Author(s)
      玉井智貴, 森田一世, 篠原美紀
    • Organizer
      第26回DNA複製・組換え・修復ワークショップ
    • Related Report
      2021 Annual Research Report
  • [Presentation] 出芽酵母における DSB修復経路での Pso2ヌクレアーゼの機能解析2021

    • Author(s)
      笠井公輔, 篠原美紀
    • Organizer
      第26回DNA複製・組換え・修復ワークショップ
    • Related Report
      2021 Annual Research Report
  • [Presentation] 細胞内エネルギー環境とDNA修復経路選択の関係性の解析2021

    • Author(s)
      辻本怜, 篠原美紀
    • Organizer
      第26回DNA複製・組換え・修復ワークショップ
    • Related Report
      2021 Annual Research Report
  • [Presentation] PP4は減数分裂期染色体軸構造形成に必要である2021

    • Author(s)
      Ke, Li, 篠原美紀
    • Organizer
      第39回 染色体ワークショップ・第19回 核ダイナミクス研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 減数分裂期で染色体数を正確に半分にするための染色体構造の役割2021

    • Author(s)
      篠原 美紀
    • Organizer
      酵母合同シンポジウム
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] 減数分裂期特異的Mek1Chk2キナーゼ 機能分離変異株の解析2021

    • Author(s)
      鈴木拓弥,浜野有希,篠原美紀
    • Organizer
      第38回染色体ワークショップ・第19回核ダイナミクス研究会
    • Related Report
      2020 Research-status Report
  • [Presentation] DNA二本鎖切断修復を誤りがちにする要因は何か?2020

    • Author(s)
      篠原美紀
    • Organizer
      第90回日本遺伝学会シンポジウム
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 減数分裂期特異的チェックポイントキナーゼMek1 Chk2/Rad53の多段階的機能2020

    • Author(s)
      鈴木拓弥,篠原美紀
    • Organizer
      第90回日本遺伝学会
    • Related Report
      2020 Research-status Report
  • [Presentation] Synaptonemal complex central regions modulates crossover pathways and feedback control of meiotic DSB formation2020

    • Author(s)
      Miki Shinohara
    • Organizer
      第43回日本分子生物学会シンポジウム「Dynamic and structural regulation of chromosome inheritance in meiosis」
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] DNA損傷チェックポイント因子の減数分裂期組換え制御における機能2019

    • Author(s)
      篠原美紀
    • Organizer
      第42回分子生物学会年会ワークショップ
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] Functional analysis of DNA damage checkpoint factors in meiotic chromosome construction2019

    • Author(s)
      Miki Shinohara
    • Organizer
      Chromosome Dynamics: An international symposium on chromatin and chromosome stability
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 減数分裂期組換え制御におけるMec1ATRと9-1-1損傷クランプの機能分担2019

    • Author(s)
      篠原美紀, Douglas K. Bishop, 篠原彰
    • Organizer
      第91回日本遺伝学会大会
    • Related Report
      2019 Research-status Report
  • [Remarks] 近畿大学アグリ技術革新研究所 研究活動報告

    • URL

      https://www.kindai.ac.jp/atiri/topics/report/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 近畿大学 農学部・大学院 農学研究科 究活動報告

    • URL

      https://www.kindai.ac.jp/agriculture/research-and-education/activity_report/advanced-bioscience/

    • Related Report
      2021 Annual Research Report

URL: 

Published: 2019-07-04   Modified: 2023-01-30  

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