Structural basis of replication restart primosome proteins involved in DnaB helicase loading

2013 Fiscal Year Final Research Report

• Project/Area Number: 23570140
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyushu University
• Principal Investigator: ABE Yoshito 九州大学, 薬学研究科(研究院), 准教授 (60315091)
• Co-Investigator(Kenkyū-buntansha): KATAYAMA Tsutomu 九州大学, 大学院薬学研究院, 教授 (70264059)
• Project Period (FY): 2011 – 2013
• Keywords: 複製再開始プライモソーム / タンパク質-DNA複合体 / タンパク質構造 / DnaT / PriC

Research Abstract

We performed the structure and function analysis of PriB, DnaT and PriC proteins engaged in replication restart in Escherichia coli. PriB is a single-stranded DNA (ssDNA) binding protein. Our NMR and FRET results showed that PriB bound to ssDNA in two-step binding manner, suggesting the cooperative binding between PriB and ssDNA. We also performed domain analysis of DnaT and PriC. From the domain information of DnaT, we suggested that the N-terminal domain of DnaT was involved in trimer formation and interaction with PriB, and the C-terminal domain of DnaT was involved in ssDNA binding based on the structure determined by NMR analysis. Furthermore, the domain information of PriC suggested that the C-terminal domain of PriC was involved in the ssDNA and SSB (single-stranded DNA binding protein) binding. Additionally, we determined the N-terminal domain of PriC using NMR analysis. Together with these results, we proposed the model of replication restart in Escherichia coli.

Research Products

• [Journal Article] Involvement of histidine in complex formation of PriB and single-stranded DNA (2014)
  • Author(s): Fujiyama, S, Abe, Y, Takenawa, T, Aramaki, T, Shioi, S, Katayama, T & Ueda, T
  • Journal Title: Biochim Biophys Acta Volume: 1844 Pages: 299-307
  • Peer Reviewed
• [Journal Article] Solution structure of the N-terminal domain of a replication restart primosome factor (2013)
  • Author(s): Aramaki, T, Abe, Y, Katayama, T & Ueda, T
  • Journal Title: PriC, in Escherichia coli. Protein Sci Volume: 22 Pages: 1279-1286
  • Peer Reviewed
• [Journal Article] Domain separation and characterization of PriC, a replication restart primosome factor in Escherichia coli (2013)
  • Author(s): Aramaki, T, Abe, Y, Ohkuri, T, Mishima, T, Yamashita, S, Katayama, T & Ueda, T
  • Journal Title: Genes Cells Volume: 18 Pages: 723-732
  • Peer Reviewed
• [Journal Article] The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA (2013)
  • Author(s): Su'etsugu, M, Harada, Y, Keyamura, K, Matsunaga, C, Kasho, K, Abe, Y, Ueda, T & Katayama, T
  • Journal Title: Environ Microbiol Volume: 15 Pages: 3183-3195
  • Peer Reviewed
• [Presentation] 大腸菌複製再開始因子PriCのSSB 結合およびDNA 結合に関する変異体解析 (2013)
  • Author(s): 荒牧峻彦, 阿部義人, 古谷香織, 片山勉, 植田正
  • Organizer: 日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20130000
• [Presentation] DNA 複製再開始に関わる大腸菌プライモソーム構成因子PriB・DnaT の相互作用解析 (2013)
  • Author(s): 藤山紗希, 阿部義人, 谷純也, 浦辺大志,片山勉, 植田正
  • Organizer: 日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20130000
• [Presentation] Domain separation and characterization of PriC, a replication restart primosome factor in Escherichia coli (2013)
  • Author(s): Takahiko Aramaki, Yoshito Abe, Takatoshi Ohkuri, Tomonori Mishima, Kaori Furutani, Shoji Yamashita, Tsutomu Katayama, Tadashi Ueda
  • Organizer: Protein Society 2013 symposium
  • Place of Presentation: USA
  • Year and Date: 20130000
• [Presentation] DNA 複製再開始に関わる大腸菌プライモソーム構成因子PriB の変異体解析 (2012)
  • Author(s): 藤山紗希, 阿部義人, 塩井誠次郎, 竹縄太一, 片山勉, 植田正
  • Organizer: 日本生化学会
  • Place of Presentation: 福岡
  • Year and Date: 20120000
• [Presentation] 大腸菌プライモソーム構成因子PriC の塩基性アミノ酸残基に関する変異体解析 (2012)
  • Author(s): 古谷佳織, 荒牧峻彦, 阿部義人, 片山勉, 植田正
  • Organizer: 日本生化学会
  • Place of Presentation: 福岡
  • Year and Date: 20120000
• [Presentation] 大腸菌プライモソーム構成因子PriC のDNA 結合に関する解析 (2012)
  • Author(s): 荒牧峻彦, 古谷佳織, 阿部義人, 片山勉, 植田正
  • Organizer: 日本生化学会
  • Place of Presentation: 福岡
  • Year and Date: 20120000
• [Presentation] 大腸菌複製再開始における分子機構の構造生物学的研究 (2012)
  • Author(s): 阿部義人
  • Organizer: 第38回日本応用酵素協会研究発表会
  • Place of Presentation: 大阪
  • Year and Date: 20120000
• [Presentation] PriC が一本鎖および二本鎖DNA いずれにも結合することの意義 (2012)
  • Author(s): 荒牧峻彦、阿部義人、古谷佳織、片山勉、植田正
  • Organizer: 日本タンパク質科学会
  • Place of Presentation: 名古屋
  • Year and Date: 20120000
• [Presentation] DNA 複製再開に関わる大腸菌プライモソーム構成因子DnaTの機能解析 (2012)
  • Author(s): 藤山紗季、阿部義人、谷純也、浦辺大志、片山勉、植田正
  • Organizer: 平成24 年度 日本生化学会九州支部例会
  • Place of Presentation: 福岡
  • Year and Date: 20120000
• [Presentation] DNA 複製再開始因子PriC の機能解析 (2011)
  • Author(s): 荒牧峻彦、阿部義人、古谷佳織、片山勉、植田正
  • Organizer: 日本タンパク質科学会
  • Place of Presentation: 大阪
  • Year and Date: 20110000
• [Remarks] PriCN 末端ドメイン構造:2RT6(DB 登録)
• [Remarks] DnaTC 末端ドメイン構造:2RU8(DB 登録)
• [Remarks] PriCN 末端ドメインNMR 帰属:11525(BMRB 登録)
• [Remarks] DnaTC 末端ドメインNMR 帰属:11549(BMRB 登録)
• [Remarks] PriB NMR 帰属:11527