The Functional Research Of Specific Proteins Interacting With 3'utr Of Erβ MRNA

ERβis a member of the nuclear receptor super family. There are researchs show that ERβis related to many malignant diseases closely and ERβplays an important role in the proliferation, metastasis and apoptosis of multiple cancers. Simultaneously ERβmay have a role in inhibiting the development of breast cancer. With recealing the interaction mechanisms of the co-regulation molecular of ERβwe can develop much safer and more effective medicines which are act on ERβspecificly. 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. The translation regulation is one of the most important way in the regulation of gene expression in eucaryote. The untranslated regions, which have important effects in translation regulation, have became one of the research hotspots in the regulation of gene expression. The 3'UTR of mRNA can control the stability, subcellular localization, degradation rate and utilization efficiency of mRNA. Thence it is significant for studying the regulation mechanisms which are related to the 3'UTR of ERβ.On the basis of MALDI-TOF-TOF we use the technology of RNA pull down to filter the special RNA binding proteins, which interact with ERβmRNA 3'UTR, from the total proteins of the breast cancer cell, MCF-7, in vitro. We identified seven proteins by MALDI-TOF-TOF MS analysis. They are vigilin, LRP130, Splicing Factor , ILF3, KHRP, SRC homolog3 and hnRNP A/B. Every protein plays special important role in differential RNA metabolic processes. Such as ILF3 has a special affinity for the mRNAs which have an AU-rich sequence. And ILF3 have the function of regulating gene expression. Vigilin, which can be induced by estrogen, is a multifunctional protein which has 14 KH domain. The expression of ERβmay be regulated by vigilin. In the following works we explore the regulation mechanism of vigilin primarily. By transcribing different fragment of ERβmRNA 3'UTR, we find out that vigilin combines to two areas of ERβmRNA 3'UTR. One fragment is from 8 bp to 207 bp and another fragment is from 294 bp to 430 bp. We thought that the controlling elements of vigilin are located in those two areas. This result is a foundantion to revealing the regulation mechanism of vigilin in regulating the expression of ERβ. The function of the other six proteins we identified in regulating ERβmRNA and the concrete regulation mechanisms of those proteins are unknown currently. And thoes issues would be solved in the follow-up works.