Identification of novel biding partners for oncogenes of Ewing sarcoma

• Project/Area Number: 16K20345
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Pediatric surgery
• Research Institution: Yamaguchi University
• Principal Investigator: KITAGAWA Takao 山口大学, 大学院医学系研究科, 助教 (20614928)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: BioID / プロテオミクス / ユーイング肉腫 / ewing sarcoma / EWS-FLI1 / 酵母 / Yeast two-hybrid / 癌 / 応用微生物 / 遺伝子 / 蛋白質 / 微生物

Outline of Final Research Achievements

Ewing sarcoma is a bone or soft tissue cancer, which is caused by chromosomal rearrangement. The resulting EWS-FLI1, EWS-ERG, EWS-E1AF show oncogenic activity, but its molecular mechanism is almost unclear. To identify binding partners for EWS-FLI1, I performed Two-hybrid screening for human cDNA and co-immunoprecipitation of EWS-FLI1. As a result, no binding partners were obtained in the two-hybrid screening, but RBM14 was obtained in the co-immunoprecipitation screening. Further, I tried BioID method, which is proximity biotination assay for target proteins. I obtained SFPQ, NONO, PSPC1, and FUS, which are part of paraspecle components, as a interaction partner. However, these proteins did not show any interaction for EWS-FLI1. Now, I have tried development of piggybac system of BioID method for EWS-FLI1.