Identification of Elongin A target genes and clarification of the mechanisms governing conversion of the Elongin complex from its elongation factor to its ubiquitin ligase form

• Project/Area Number: 15K08279
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kochi University
• Principal Investigator: Aso Teijiro 高知大学, 教育研究部医療学系基礎医学部門, 教授 (20291289)
• Co-Investigator(Renkei-kenkyūsha): KITAJIMA Shigetaka 東京医科歯科大学, 難治疾患研究所, 教授 (30186241)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: Elongin / CSB / ユビキチンリガーゼ / 転写伸長 / RNAポリメラーゼII / DNA傷害 / ストレス / 遺伝子発現 / 胚様体 / レチノイン酸 / FRET / small nuclear RNA / non-coding RNA

Outline of Final Research Achievements

Elongin A (EloA) performs dual functions as the transcriptionally active subunit of RNA polymerase II (Pol II) elongation factor Elongin and as the substrate recognition subunit of an ubiquitin ligase (E3) that ubiquitylates Pol II in response to DNA damage. In this study, we first identified Hox genes as the targets of elongation factor EloA. We also investigated the mechanisms governing conversion of the Elongin complex from its elongation factor to its E3 form. Assembly of EloA into the E3 was strongly induced by DNA-damaging agents; and α-amanitin, a drug that induces Pol II stalling; and by other various stimuli. In addition, we demonstrated (i) that EloA and the E3 subunit Cul5 associate in cells with the Cockayne syndrome B (CSB) protein, (ii) that this interaction was also induced by DNA-damaging agents and α-amanitin, and (iii) that CSB protein promotes stable recruitment of the EloA-E3 to sites of DNA damage.

Research Products

• [Int'l Joint Research] Stowers Institute for Medical Research(米国)
• [Int'l Joint Research] Stowers Institute for Medical Research(米国)
• [Int'l Joint Research] Stowers Institute for Medical Research/University of Kansas Medical Center(米国)
• [Journal Article] Cockayne syndrome B protein regulates recruitment of the Elongin A ubiquitin ligase to sites of DNA damage. (2017)
  • Author(s): Weems JC, Slaughter BD, Unruh JR, Boeing S, Hall SM, McLaird MB, Yasukawa T, Aso T, Svejstrup JQ, Conaway JW, Conaway RC
  • Journal Title: J. Biol. Chem. Volume: 292 Issue: 16 Pages: 6431-6437
  • DOI: 10.1074/jbc.c117.777946
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Assembly of the Elongin A ubiquitin ligase is regulated by genotoxic and other stresses. (2015)
  • Author(s): Weems J.C., Slaughter B.D., Unruh J.R., Hall S.M., McLaird M.B., Gilmore J.M., Washburn M.P., Florens L., Yasukawa, T., Aso, T., Conaway, J.W., and Conaway, R.C.
  • Journal Title: J. Biol. Chem. Volume: 290 Issue: 24 Pages: 15030-15041
  • DOI: 10.1074/jbc.m114.632794
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] ChIP-sequence解析を用いた転写伸長因子Elongin Aの標的遺伝子の同定 (2016)
  • Author(s): 安川孝史、筒井文、佐藤チエリ、佐藤滋生、Ronald C. Conaway、Joan W. Conaway、麻生悌二郎
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-12-02