SUMOylation of Grb2 enhances the ERK activity by increasing its binding with Sos1

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• 作者：Yingying Qu (6)
  Qin Chen (6)
  Xueping Lai (6)
  Changhong Zhu (6)
  Cheng Chen (6)
  Xian Zhao (6)
  Rong Deng (6)
  Ming Xu (6)
  Haihua Yuan (6) (8)
  Yanli Wang (6)
  Jianxiu Yu (6) (7) (8)
  Jian Huang (6)
  
  6. Department of Biochemistry and Molecular Cell Biology ; Shanghai Key Laboratory of Tumor Microenvironment and Inflammation ; Shanghai Jiao Tong University School of Medicine (SJTU-SM) ; Shanghai ; 200025 ; China
  8. Department of Oncology ; No. 3 People鈥檚 Hospital Affiliated to Shanghai Jiao Tong University School of Medicine ; Shanghai ; China
  7. State Key Laboratory of Oncogenes and Related Genes ; Shanghai Jiao Tong University School of Medicine ; Shanghai ; China
  
• 关键词：Grb2 ; SUMOylation ; Sos1 ; ERK activity ; Tumorigenesis ; Cell migration
• 刊名：Molecular Cancer
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：13
• 期：1
• 全文大小：1,607 KB
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• 刊物主题：Cancer Research; Oncology;
• 出版者：BioMed Central
• ISSN：1476-4598

文摘

Background Grb2 (Growth factor receptor-bound protein 2) is a key adaptor protein in maintaining the ERK activity via linking Sos1 (Son of sevenless homolog 1) or other proteins to activated RTKs, such as EGFR. Currently, little knowledge is available concerning the post-translational modification (PTM) of Grb2 except for its phosphorylation. Since emerging evidences have highlighted the importance of SUMOylation (Small ubiquitin-related modifier), a reversible PTM, in modulating protein functions, we wondered if Grb2 could be SUMOylated and thereby influences its functions especially involved in the Ras/MEK/ERK pathway. Methods SUMOylation of Grb2 was analyzed with the in vivo SUMOylation assay using the Ni2+-NTA affinity pulldown and the in vitro E.coli-based SUMOylation assay. To test the ERK activity and cell transformation, the murine fibroblast cell line NIH/3T3 and the murine colon cancer cell line CMT-93 were used for the experiments including Grb2 knockdown, ectopic re-expression, cell transformation and migration. Immunoprecipitation (IP) was employed for seeking proteins that interact with SUMO modified Grb2. Xenograft tumor model in mice was conducted to verify that Grb2 SUMOylation regulated tumorigenesis in vivo. Results Grb2 can be SUMOylated by SUMO1 at lysine 56 (K56), which is located in the linker region between the N-terminal SH3 domain and the SH2 domain. Knockdown of Grb2 reduced the ERK activity and suppressed cell motility and tumorigenesis in vitro and in vivo, which were all rescued by stable ectopic re-expression of wild-type Grb2 but not the mutant Grb2K56R. Furthermore, Grb2 SUMOylation at K56 increased the formation of Grb2-Sos1 complex, which sequentially leads to the activation of Ras/MEK/MAPK pathway. Conclusions Our results provide evidences that Grb2 is SUMOylated in vivo and this modification enhances ERK activities via increasing the formation of Grb2-Sos1 complex, and may consequently promote cell motility, transformation and tumorigenesis.