Identification Of RBM47 ISGylation And Preliminary Exploration Of Its Function

Cancer is the number one disease that threatens the life and health of mankind.RBM47 is an RNA binding protein with three RNA recognition domains,which can inhibit the growth,proliferation,invasion and migration of cancer cells.HERC5,the E3 ligase of ISGylation modified by a ubiquitin-like protein,was discovered by analyzing the mass spectrometry data of intracellular overexpression of RBM47 and enrichment of its interacting proteins.Using si RNA to specifically knock down HERC5 and ISG15,we found that RBM47 protein dimerization was reduced,indicating that RBM47 may have ISGylation modification.By constructing the ISGylation modification system,overexpression of ISGylation modification related proteins in the cell,we further demonstrated that ISGylation can occur on RBM47.Subsequently,lysine 329 was identified as the modification site,and it was found that the modification site promoted the dimerization of RBM47 forming proteins.In looking for the upstream pathway of ISGylation affecting RBM47,we found that the hypoxia environment could promote the modification.In addition,the multipoint phosphorylation of RBM47 itself also promotes the occurrence of modification.We demonstrated that RBM47 interacts with YTHDF2 by yeast double hybridization,and found that RBM47 no longer interacts with YTHDF2 after the glycine mutation at position 538 to arginine.When we explored the function of RBM47 ISGylation modification,we used the isg15-induced low si RNA experiment combined with the intracellular immunoprecipitation experiment of RBM47,and found that ISGylation modification of RBM47 could weaken the protein interaction between RBM47 and YTHDF2.In general,this paper is to study the ISG mechanism of RBM47 and explore the function of this modification.