Interaction Between Wheat EEF1B(TaeEF1B) And Chinese Wheat Mosaic Virus(CWMV) And Its Effect On The Viral Replication

Eukaryotic translation elongation factors(e EFs)are multi-function regulated proteins.e EFs can interact with a variety of viral proteins,viral RNAs,or untranslated regions of the genome(UTR)and are conducive to viral life cycles.The interaction between e EF1 B,an important component of e EF,and virus remains to be explored.Chinese wheat mosaic virus(CWMV),transmitted by Polymyxa graminis,has been identified to be a new virus on winter wheat plants in Shandong Province,China,in our lab and significant progresses have been made in its molecular biology.However,the interaction between CWMV and host factors remains largely unknown.In this study,we first cloned the gene of wheat e EF1 B and analyzed its homology,expression pattern,interaction with CWMV.The following results are obtained:In this study,eEF1 B of wheat was cloned by RT-PCR and named TaeEF1 B.Homologous analysis suggested that Tae EF1 B was highly conserved and the region 137-226 aa was its conserved domain.Gene expression of Tae EF1 B analysis showed Tae EF1 B was expressed on the highest m RNA level in stem.In addition,we constructed the prokaryotic expression vector and gained its purified protein by induced expression,which laid foundation of studying its role in CWMV infection.Then,the relationship between CWMV and e EF1 B was analyzed by CWMV infection clone in plant Nicotiana benthamiana(Nb).It was found by q PCR and Northern blotting that the gene silencing of Nbe EF1 B in Nb inhibited the accumulation of CWMV CP and genome RNAs.Otherwise,the transient overexpression of Nbe EF1? in Nb promoted the accumulation of CWMV CP and genome RNAs,suggesting that Nbe EF1 B could regulate CWMV replication positively.The interaction between Tae EF1 B and CWMV was analyzed by yeast two-hybrid(Y2H)and Electrophoretic Mobility Shift Assay(EMSA).Yeast two-hybrid assays suggested that Tae EF1 B could not interact with CWMV Rd Rp,while EMSA analysis showed that it had the nucleotide-protein interaction between CWMV genome and Tae EF1 B,and its binding site was only in the 3'UTR of the CWMV positive-sense strand,further illustrating that Tae EF1 B could promote viral replication by specifically binding the viral genomic 3'UTR.These results could lay foundation for further investigation of functional mechanism that Tae EF1 B could promotes CWMV replication.