Identification of factors involved in the eukaryotic mismatch repair reaction

• Project/Area Number: 25650011
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: Osaka University
• Principal Investigator: TAKAHASHI Tatsuro 大阪大学, 理学(系)研究科(研究院), 助教 (50452420)
• Research Collaborator: KAWASOE Yoshitaka ; TERUI Riki
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: ミスマッチ修復 / クロマチン / DNA損傷応答 / DNA複製 / ツメガエル / 卵抽出液 / 試験管内系 / DNA修復 / 染色体 / 無細胞系

Outline of Final Research Achievements

Eukaryotic chromosomal DNA is wrapped around histones to be assembled into chromatin, whose dynamic remodeling is linked to various DNA transactions. The mismatch repair system maintains DNA replication fidelity by correcting replication errors. Impairment of this system thus leads to oncogenesis in human. Here, we sought factors involved in the mismatch repair reaction in the context of chromatin, using Xenopus egg extract that promotes chromatin assembly in vitro. We have identified at least 9 factors whose involvements in mismatch repair have not previously been documented, and 3 of which were chromatin-related factors. Having identified these new factors, this work established an important step toward understanding of the mismatch repair reaction on chromatin.

Research Products

• [Journal Article] RecQ4 promotes the conversion of the pre-initiation complex at a site-specific origin for DNA unwinding in Xenopus egg extracts (2015)
  • Author(s): Sanuki, Kubota, Kanemaki MT, Takahashi TS, Mimura S, Takisawa H
  • Journal Title: Cell Cycle Volume: 14-7 Issue: 7 Pages: 1010-23
  • DOI: 10.1080/15384101.2015.1007003
  • Peer Reviewed
• [Journal Article] Thymine DNA glycosylase is a CRL4Cdt2 substrate. (2014)
  • Author(s): Slenn TJ, Morris B, Havens CG, Freeman RM Jr, Takahashi TS, Walter JC.
  • Journal Title: The Journal of Biological Chemistry Volume: 289 Issue: 33 Pages: 23043-55
  • DOI: 10.1074/jbc.m114.574194
  • Peer Reviewed
• [Journal Article] MutS stimulates the endonuclease activity of MutL in an ATP-hydrolysis-dependent manner. (2013)
  • Author(s): Shimada, A., Kawasoe, Y., Hata, Y., Takahashi, T.S., Masui, R., Kuramitsu, S., Fukui, K.
  • Journal Title: FEBS Journal Volume: 280 (14) Issue: 14 Pages: 3467-3479
  • DOI: 10.1111/febs.12344
  • Peer Reviewed
• [Presentation] PCNAとXEco2はDNA複製と協調したSororin染色体結合反応に機能する (2014)
  • Author(s): 高橋 達郎
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] ミスマッチ修復因子MutSαはPCNAのDNAからの解離を阻害することで新生鎖の記憶を保持する (2014)
  • Author(s): 河添 好孝
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] MutSα依存的なミスマッチ塩基対周辺のヌクレオソーム排除反応 (2014)
  • Author(s): 照井 利輝
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] MutSα-dependent exclusion of nucleosomes from mismatch sites in Xenopus egg extracts (2014)
  • Author(s): 高橋 達郎
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 静岡県御殿場市 ホテル時之栖
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] PCNA and XEco2 couple chromatin loading of Sororin to DNA replication in Xenopus egg extracts (2014)
  • Author(s): 東 寅彦
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 静岡県御殿場市 ホテル時之栖
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] MutSα prevents dissociation of PCNA from DNA to keep strand information for eukaryotic mismatch repair (2014)
  • Author(s): 河添 好孝
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 静岡県御殿場市 ホテル時之栖
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] MutSα prevents PCNA unloading to keep strand information for eukaryotic mismatch repair (2014)
  • Author(s): 高橋 達郎
  • Organizer: 国立遺伝学研究所・研究集会「染色体 DNA の安定維持の分子メカニズム」
  • Place of Presentation: 静岡県三島市 国立遺伝学研究所
  • Year and Date: 2014-11-06 – 2014-11-07
  • Invited
• [Presentation] MutSα prevents dissociation of PCNA from DNA to keep strand information for eukaryotic mismatch repair (2014)
  • Author(s): 高橋 達郎
  • Organizer: The 5th US-Japan DNA repair meeting
  • Place of Presentation: 徳島県鳴門市 グランドエクシブ鳴門
  • Year and Date: 2014-10-28 – 2014-10-31
• [Presentation] ミスマッチ修復とクロマチン複製のカップリング (2014)
  • Author(s): 高橋 達郎
  • Organizer: 蛋白研セミナー 『染色体伝承の分子背景：複製から染色体分離まで』
  • Place of Presentation: 大阪府吹田市 大阪大学蛋白質研究所 1 階講堂
  • Year and Date: 2014-09-25 – 2014-09-26
  • Invited
• [Presentation] MutSα-dependent eviction of nucleosomes around mismatch sites in Xenopus egg extracts (2014)
  • Author(s): Tatsuro Takahashi, Riki Terui, Kanae Taki, koji Nagao, Takuro Nakagawa, Yumiko Kubota, Haruhiko Takisawa, Chikashi Obuse, Hisao Masukata
  • Organizer: International Conference: Replication, repair, and transcription; coupling mechanisms and chromatin dynamics for genome integrity
  • Place of Presentation: 京都市左京区 京都大学 時計台100周年記念ホール
• [Presentation] 真核生物ミスマッチ修復機構とクロマチン動態、DNA損傷応答機構のクロストーク (2014)
  • Author(s): 高橋達郎
  • Organizer: 平成25年度「がん研究分野の特性等を踏まえた支援活動」 公開シンポジウム
  • Place of Presentation: 東京都千代田区 学術総合センター
  • Invited
• [Presentation] Essential roles of the N-terminal region of XEco2 acetyltransferase for cohesion establishment in Xenopus egg extracts (2013)
  • Author(s): Torahiko Higashi, Takuro Nakagawa, Hisao Masukata, Tatsuro Takahashi
  • Organizer: Cold Spring Harbor Laboratory Meeting "Eukaryotic DNA replication & Genome Maintenance"
  • Place of Presentation: Cold Spring Harbor, NY
• [Presentation] MutSα-dependent eviction of nucleosomes around mismatch sites in Xenopus egg extracts (2013)
  • Author(s): Riki Terui, Kanae Taki, Torahiko Higashi, Takuro Nakagawa, Hisao Masukata, Tatsuro Takahashi
  • Organizer: Cold Spring Harbor Laboratory Meeting "Eukaryotic DNA replication & Genome Maintenance"
  • Place of Presentation: Cold Spring Harbor, NY
• [Presentation] Evidence that DNA-loaded PCNA acts as the strand marker for eukaryotic mismatch repair (2013)
  • Author(s): Yoshitaka Kawasoe, Toshiki Tsurimoto,Takuro Nakagawa, Hisao Masukata, Tatsuro Takahashi
  • Organizer: Cold Spring Harbor Laboratory Meeting "Eukaryotic DNA replication & Genome Maintenance"
  • Place of Presentation: Cold Spring Harbor, NY
• [Presentation] MutSα-dependent eviction of nucleosomes around mismatch sites in Xenopus egg extracts (2013)
  • Author(s): Tatsuro Takahashi, Riki Terui, Kanae Taki, Koji Nagao, Takuro Nakagawa, Uimiko Kubota, Haruhiko Takisawa, Chikashi Obuse, Hisao Masukata
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 兵庫県神戸市 神戸国際会議場
  • Invited
• [Presentation] ミスマッチ周辺でのMutS依存的ヌクレオソーム除去反応に関わる因子の探索 (2013)
  • Author(s): Riki Terui, Kanae Taki, koji Nagao, Takuro Nakagawa, Yumiko Kubota, Haruhiko Takisawa, Chikashi Obuse, Hisao Masukata
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 兵庫県神戸市 神戸国際会議場
• [Presentation] ミスマッチ修復機構がクロマチン複製の場で機能するための機構 (2013)
  • Author(s): 高橋達郎
  • Organizer: 国立遺伝学研究所・研究会「染色体 DNA の安定維持の分子メカニズム」
  • Place of Presentation: 静岡県三島市 国立遺伝学研究所
• [Presentation] ミスマッチ修復機構がクロマチン複製の場で機能するための機構 (2013)
  • Author(s): 高橋達郎
  • Organizer: 国立遺伝学研究所・研究会「クロマチンによる遺伝情報のエピジェネティック制御機構」
  • Place of Presentation: 静岡県三島市 国立遺伝学研究所
• [Presentation] Coupling of cohesin acetylation to the process of DNA replication in Xenopus egg extracts
  • Author(s): Torahiko Higashi, Takuro Nakagawa, Hisao Masukata, Tatsuro Takahashi
  • Organizer: The 18th IMCB Symposium
  • Place of Presentation: 東京都文京区 東京大学弥生講堂
  • Invited