Research Of Specific Proteins Interacting With3’UTR Of GW112mRNA

Objective Human olfactomedin4(OLFM4) gene as known as the anti-apoptotic molecule GW112is found to be frequently up-regulated in many types of human tumors and it was believed to play significant role in the progression of gastric cancer. In eucaryote most controlling elements of mRNA are located in the untranslated regions. By integrating with specific RNA binding protein those controlling elements can play important roles in regulating the translation of mRNA. This study aims to filter the special RNA binding proteins which interact with GW112mRNA3’UTR and to analyze the character of protein.Methods By transcribing and marking the fragment of GW112mRNA3’UTR,we used the technology of magnetic beads adsorption to filter the GW112mRNA3’UTR binding proteins, from the total proteins of the gastric cancer cell, SGC-7901, in vitro. We identified proteins by mass spectrometry and western blot.Results Here,we describe the affinity purification protein (100-120kd) that binds to GW112mRNA3’UTR, Mass spectrometry revealed that the identity of the protein was nucleolin. After detection with anti-nucleolin, the target band appeared on SDS-PAGE,which correspond with the MS results.Conclusion Our result demonstrated that nucleolin could specifically binding to GW112mRNA3’UTR, which maybe influence the stability of G W112m RN A，a nd t hi s re s ul t w il l pr ov i d e go o d ba si c fo r fu n c ti o na lre se a rc h of G W112i n g a st r i c c a nc er c el l a po p t o s is.