BmArrestin2 Gene Function In Silkwom,Bombyx Mori

Beta arrestin2 is one of the widely distributed protein,in mammals,it plays an important regulatory role in the desensitization,internalization and sensitive reaction of the G protein coupled receptor?G protein-coupled receptors,GPCRs?,it is also very important to cell proliferation and has the important status in gene transcription.?eta arrestin2 is highly conserved with homologs in many metazoans from Drosophila to mice to humans.Now,in this pattern of lepidoptera insects of the silkworm,the function of the beta arrestin2 has never been reported so far.This research obtained BmArrestin2gene,detected expression level of these genes in different tissues and diverse developmental stages of the silkworm,investigated the impact of overexpression BmArrestin2 on the size,cycle and apoptosis of cultured cells as well as part of signaling pathways genes in silkworm.Discussed the BmArrestin2 gene effects on virus infected cells.1.The the expression profiling of the Bm Arrestin2 gene of silkworm,subcellular localization and the interaction protein of BmArrestin2In order to investigate the expression of BmArrestin2 gene,real-time quantitative PCR was performed to analysis the expression levels of BmArrestin2 gene in gonads in diverse developmental stages and in diverse tissues in the 3^rdday of 5^thinstar.BmArrestin2 in the ovarian tissue of 3^rdday of 5^thwere the highest expression levels,in other diverse developmental stages were in a low volume.The abundance of BmArrestin2 mRNA in the tracheal was the highest,was lower in the testis.In order to investigate the cellular localization features of BmArrestin2,BmArrestin2 protein antibodies were prepared.Immunofluorescence showed that BmArrestinr2 in ovarian sources of BmN cells can express,and are mainly distributed in cytoplasm.in order to further study the function of BmArrestin2.CO-Immunoprecipitation was performed to detect the interaction protein of BmArrestin2,the mass spectrum identification results suggest Actin and Yorkie3?Yki3?may be the interaction proteins.2.The effects on the cultured cells of silkworm after regulation of expression on BmArrestin2 geneIn order to research the function of BmArrestin2 gene,we selected some signal pathway genes for quantitative detection.The results showed that when overexpression BmArrestin2 gene,the expression level of Hippo pathway kibra,ex genes significantly decreased compared with control cells;wts,Bmpr gene have no changes;wnt,yki,cyce,iap,Fj,dpp,serr,myc,crb gene expression level significantly increased.In the genetic pathways associated with growth,GASP,NkAB,DYNE,PSLR expression level significantly increased;JH,PCL,LSAM gene expression quantity has no obvious change;While ECDY,CFAP,LACH,UMX significantly decreased compared with control cells.In natural immune and apoptosis related pathways,STAT,Tal,SPZ,APA,PGRP-LB,PGRP-LE genes expression levels decreased while there was no significant alteration of the SURF,QCTY gene expression levels.After BmArrestin2gene silence,some genes expression levels were contrary to the results when overexpression.The volume and the cell cycle of cultured cells were decreased compared with control BmN cells,bue have no obvious affect on apoptosis.3.BmArrestin2 effect on virus infected cellsIn order to further identify BmArrestin2 gene function,we detect the BmArrestin2effect on virus infected cells.The results showed that overexpression BmArrestin2 cells can enhance the infection of BmNPV and BmCPV to cells.In order to further explore this mechanism,antibodies was add to cells in order to blocking endogenous BmArrestin2,and the results showed after blocking BmArrestin2,the infection of BmNPV to cells was decreased.We also find that the expression level of endogenous BmArrestin2 can decreased when infected BmNPV and BmCPV,we speculated that this phenomenon may be caused by the BmArrestin2 regulation to the inherent cell immune.