Structural and functional analysis of the mechanism underlying perturbation of Csk by Helicobacter pylori virulence factor CagA

• Project/Area Number: 26860284
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Bacteriology (including mycology)
• Research Institution: The University of Tokyo
• Principal Investigator: Hayashi Takeru 東京大学, 医学(系)研究科(研究院), 助教 (10722209)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: ピロリ菌 / CagA / 細胞内シグナル撹乱 / 構造-機能解析 / ピロリ菌感染 / タンパク質複合体構造解析 / タンパク質複合体機能解析

Outline of Final Research Achievements

We established the experimental method for expression and purification of tyrosine-phosphorylated CagA oncoprotein from Helicobacter pylori by employing bacterial expression system. Using the purified tyrosine-phosphorylated CagA protein, we elucidated that the interaction of CagA with Csk, one of the intracellular targets of CagA, is due to direct binding and that the CagA-bound Csk is aberrantly activated. Among multiple tyrosine-phosphorylation motifs in a single CagA molecule, we identified the key motif for the interaction of CagA with Csk. Furthermore, crystal structure analysis revealed that the interaction mode of CagA-Csk complex.

Research Products

• [Journal Article] Determination of the catalytic activity of LEOPARD syndrome- associated SHP2 mutants toward parafibromin, a bona fide SHP2 substrate involved in Wnt signaling (2016)
  • Author(s): Saori Noda, Atsushi Takahashi, Takeru Hayashi, Sei-ichi Tanuma and Masanori Hatakeyama
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 469 Issue: 4 Pages: 1133-1139
  • DOI: 10.1016/j.bbrc.2015.12.117
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Improvement of Helicobacter pylori CagA crystals by the multi-step soaking method (2016)
  • Author(s): Senda, M.
  • Journal Title: Cryst. Growth Des. Volume: 16 Issue: 3 Pages: 1565-1571
  • DOI: 10.1021/acs.cgd.5b01692
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Host SHP1 phosphatase antagonizes Helicobacter pylori CagA and can be downregulated by EBV. (2016)
  • Author(s): Saju, P., Murata-Kamiya, N., Hayashi, T., Senda, Y., Nagase, L., Noda, S., Matusaka, K., Funata, S., Kunita, A., Urabe, M., Seto, Y., Fukayama, M., Kaneda, A., Hatakeyama, M.
  • Journal Title: Nature Microbiology Volume: 1 Issue: 4 Pages: 16026-16026
  • DOI: 10.1038/nmicrobiol.2016.26
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Dramatic increase in SHP2 binding activity of Helicobacter pylori Western CagA by EPIYA-C duplication: its implications in gastric carcinogenesis (2015)
  • Author(s): Nagase, L.
  • Journal Title: Sci. Rep. Volume: 5 Issue: 1 Pages: 15749-15749
  • DOI: 10.1038/srep15749
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] ピロリ菌CagAによる宿主分子撹乱機構の構造と機能 (2014)
  • Author(s): 林剛瑠、長瀬里沙、千田美紀、千田俊哉、畠山昌則
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
  • Invited
• [Presentation] ピロリ菌がんタンパク質CagAと標的分子Cskとの相互作用における生物物理化学的解析 (2014)
  • Author(s): 林剛瑠、畠山昌則
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Book] 実験医学 増刊 (2014)
  • Author(s): 千田俊哉、千田美紀、林剛瑠、畠山昌則
  • Total Pages: 5
  • Publisher: 羊土社