Mechanism of intra-nuclear localization of fission yeast chromosome

• Project/Area Number: 15K14450
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: Osaka University
• Principal Investigator: Masukata Hisao 大阪大学, その他部局等, 名誉教授 (00199689)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: テロメア / 複製開始点 / 複製タイミング / 分裂酵母 / shelterin / 細胞核 / Taz1 / 細胞周期 / 複製タイミング制御 / 核内局在 / LacI-GFP

Outline of Final Research Achievements

For faithful duplication of genetic information, DNA replication initiates from many distinct sites on eukaryotic chromosomes in a manner to be regulated by a spatiotemporal program, although the molecular mechanisms of regulation has not been uncovered. We discovered that the higher-order chromatin communication of the telomeres and internal late replication origin plays important role in replication-timing control. Using LacI-GFP fluorescent protein bound to lacO repeat inserted nearby replication origins in fission yeast Shizosaccharomyces pombe, we found that late replication origins are localized at the nuclear periphery and adjacent to telomeres especially in G1/S phase when replication-timing is determined. Telomere-binding protein complex “shelterin” is required for telomere-association of the internal replication origins, although the nuclear peripheral localization is not important. These finding provide insight into a novel mechanism of replication control by telomeres.

Academic Significance and Societal Importance of the Research Achievements

これまで複製開始制御と核内配置の関係は明らかにされていなかったが、目印となる蛍光タンパク質を特定の複製開始点に結合させることにより、生きた酵母細胞で複製開始点の高解像度観察が可能になった。本研究は、染色体末端テロメアが遠く離れた染色体内部とコミュニケーションするしくみを明らかにしたことになり、将来テロメアが遠隔制御する疾患原因遺伝子の発見ならびに疾患の予防につながる可能性がある。

Research Products

• [Journal Article] Nucleosomes around a mismatched base pair are excluded via an Msh2-dependent reaction with the aid of SNF2 family ATPase Smarcad1. (2018)
  • Author(s): Terui R, Nagao K, Kawasoe Y, Taki K, Higashi TL, Tanaka S, Nakagawa T, Obuse C, Masukata H, Takahashi TS
  • Journal Title: Genes & development Volume: 32(11-12) Issue: 11-12 Pages: 806-821
  • DOI: 10.1101/gad.310995.117
  • Peer Reviewed / Open Access
• [Journal Article] Shelterin promotes tethering of late replication origins to telomeres for replication‐timing control (2018)
  • Author(s): Ogawa Shiho、Kido Sayuri、Handa Tetsuya、Ogawa Hidesato、Asakawa Haruhiko、Takahashi Tatsuro S、Nakagawa Takuro、Hiraoka Yasushi、Masukata Hisao
  • Journal Title: The EMBO Journal Volume: 37 Issue: 15 Pages: 15-15
  • DOI: 10.15252/embj.201898997
  • Peer Reviewed / Open Access
• [Journal Article] Nucleosomes around a mismatched base pair are excluded via an Msh2-dependent reaction with the aid of SNF2-family ATPase Smarcad1. (2018)
  • Author(s): Terui,R.,Nagao,K.,Kawasoe, Y., Taki,K., Higashi,TL., Tanaka,S., Nakagawa,T., Obusae,C., Masukata,H., Takahashi,TS.
  • Journal Title: Genes and Development Volume: -
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulation of mitotic recombination between DNA repeats in centromeres. (2017)
  • Author(s): Zafar,F., Okita,AK. Onaka,AT., Su,J., Katahira,Y., Nakayama,J. Takahashi,TS., Masukata, H., Nakagawa, T.
  • Journal Title: Nucleic Acids Research Volume: 44(22) Issue: 19 Pages: 11222-11235
  • DOI: 10.1093/nar/gkx763
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MutSα maintains the mismatch repair capability by inhibiting PCNA unloading (2016)
  • Author(s): Kawasoe, Y., Tsurimoto, T., Nakagawa, T., Masukata, H., Takahashi, TS.
  • Journal Title: eLIFE Volume: 12;5
  • DOI: 10.7554/elife.15155
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Rad51 and Rad54 promote noncrossover recombination between centromere repeats on the same chromatid to prevent isochromosome formation. (2016)
  • Author(s): Onaka, AT., Toyofuku, N., Inoue, T., Okita, AK., Sagawa, M., Su, J., Shitanda, T., Matsuyama, R., Zafar, F., Takahashi, TS., Masukata, H., Nakagawa, T.
  • Journal Title: Nucleic Acids Research Volume: 44 (22) Issue: 22 Pages: 10744-10757
  • DOI: 10.1093/nar/gkw874
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Shugoshin forms a specialized chromatin domain at subtelomeres that regulates transcription and replication timing. (2016)
  • Author(s): Tashiro S, Handa T, Matsuda A, Ban T, Takigawa T, Miyasato K, Ishii K, Kugou K, Ohta K, Hiraoka Y, Masukata H, Kanoh J.
  • Journal Title: Nature Commun. Volume: 7 Issue: 1 Pages: 10393-10393
  • DOI: 10.1038/ncomms10393
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Histone H4 acetylation required for chromatin decompaction during DNA replication (2015)
  • Author(s): Ruan K, Yamamoto TG, Asakawa H, Chikashige Y, Kimura H, Masukata H, Haraguchi T, Hiraoka Y.
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 1-10
  • DOI: 10.1038/srep12720
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Meiotic nuclear movements in fission yeast are regulated by the transcription factor Mei4 downstream of a Cds1-dependent replication checkpoint pathway. (2014)
  • Author(s): Ruan K, Yamamoto TG, Asakawa H, Chikashige Y, Masukata H, Haraguchi T, Hiraoka Y.
  • Journal Title: Genes Cells Volume: 20 Issue: 3 Pages: 160-172
  • DOI: 10.1111/gtc.12207
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Spatial and temporal control of chromosomal DNA replication by telomere sequence and telomere binding proteins in the fission yeast nucleus. (2017)
  • Author(s): 升方久夫
  • Organizer: The 9th International Fission Yeast Meeting (Banff, Canada)
  • Int'l Joint Research
• [Presentation] Spatial and temporal control of chromosome DNA replication by telomere binding proteins. (2017)
  • Author(s): 升方久夫
  • Organizer: Eukaryotic DNA Replication and Genome Maintenance（Cold Spring Harbor, NY USA)
  • Int'l Joint Research
• [Presentation] テロメア結合タンパク質およびシェルタリン構成因子によるDNA複製の時空間的制御機構の解明 (2017)
  • Author(s): 小川志帆、半田哲也、中村優太、高橋達郎、中川拓郎、小川英知、淺川東彦、平岡泰、升方久夫
  • Organizer: 第40回日本分子生物学会年会（神戸)
• [Presentation] 分裂酵母シェルタリンによる複製開始タイミング制御機構 (2017)
  • Author(s): 城戸沙友里、小川志帆、高橋達郎、中川拓郎、升方久夫
  • Organizer: 第40回日本分子生物学会年会（神戸)
• [Presentation] 分裂酵母における複製開始複合体形成の核内局在による制御 (2016)
  • Author(s): 升方久夫
  • Organizer: 第38回日本分子生物学会年会ワークショップ
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-11-30
  • Invited
• [Presentation] Control of DNA replication timing and location by telomere-binding proteins in fission yeast. (2016)
  • Author(s): 升方久夫
  • Organizer: 第１０回3R Symposium
  • Place of Presentation: 松江
  • Year and Date: 2016-11-13
  • Int'l Joint Research / Invited
• [Presentation] Control of subnuclear localization and replication timing by telomere binding proteins in fission yeast (2016)
  • Author(s): 升方久夫
  • Organizer: FASEB Science Research Conferences, Yeast Chromosome Structure, Replication and Segregation
  • Place of Presentation: Snowmass, Colorado, USA
  • Year and Date: 2016-07-31
  • Int'l Joint Research / Invited
• [Presentation] Control of replication timing and subnuclear localization by telomere binding proteins in fission yeast (2016)
  • Author(s): 升方久夫
  • Organizer: DNA Metabolism, Genomic Stability and Disease: Cold Spring Harbor Asia
  • Place of Presentation: Suzhou、China
  • Year and Date: 2016-06-13
  • Int'l Joint Research / Invited
• [Presentation] テロメア結合タンパク質によるDNA複製の時空間的制御機構 (2015)
  • Author(s): 小川志帆、中村優太、高橋達郎、中川拓郎、小川英知、淺川東彦、平岡泰、升方久夫
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] Replication timing control by telomere-adjacent nuclear localization. (2015)
  • Author(s): 小川志帆、中村優太、半田哲也、小川英知、淺川東彦、高橋達郎、中川拓郎、平岡泰、升方久夫
  • Organizer: Eukaryotic DNA Replication and Genome Maintenance
  • Place of Presentation: Cold Spring Harbor, NY USA
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] The mechanism of the MutSα-dependent strand memory reaction in Xenopus egg extract. (2015)
  • Author(s): 高橋達郎、河添好孝、釣本敏樹、中川拓郎、升方久夫
  • Organizer: Eukaryotic DNA Replication and Genome Maintenance
  • Place of Presentation: Cold Spring Harbor, NY USA
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Replication timing control by telomere-adjacent nuclear localization in fission yeast. (2015)
  • Author(s): 升方久夫、中村優太、小川英知、淺川東彦、高橋達郎、中川拓郎、平岡泰、小川志帆
  • Organizer: 第８回International Fission Yeast Meeting
  • Place of Presentation: 神戸
  • Year and Date: 2015-06-21
  • Int'l Joint Research / Invited
• [Presentation] Temporal and spatial regulation of replication in fission yeast. (2015)
  • Author(s): 小川志帆、中村優太、高橋達郎、中川拓郎、小川英知、淺川東彦、平岡泰、升方久夫
  • Organizer: 第８回International Fission Yeast Meeting
  • Place of Presentation: 神戸
  • Year and Date: 2015-06-21
  • Int'l Joint Research
• [Remarks] 升方研究室
  • URL: http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/masukata/
• [Remarks] テロメアに結合するタンパク質が染色体複製開始点をテロメアに引き寄せて制御することを発見
  • URL: http://www.bio.sci.osaka-u.ac.jp/newinfo/info93.html
• [Remarks] 大阪大学大学院理学研究科生物科学専攻分子遺伝学研究室ホームページ
  • URL: http://www.bio.sci.osaka-u.ac.jp/dbs01/re-paper-temp.php?id=14
• [Remarks] 升方研究室ホームページ
  • URL: http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/masukata/thesis/index.html
• [Remarks] 大阪大学大学院理学研究科生物科学専攻分子遺伝学研究室
  • URL: http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/masukata/