【摘要】 AAV-2Rep78蛋白是涉及到病毒转录、复制和位点特异性整合的多功能蛋白,它同时还介导AAV-2与其协助病毒之间的相互作用.我们通过免疫共沉淀实验证明细胞内PML蛋白与Rep78蛋白存在相互作用,作用区域位于Rep78蛋白C末端545FPCRQCERM553位置,在细胞内短暂表达Rep78会加速PML蛋白的降解.分别使用含有ICP0的病毒株G207和ICP0突变的病毒株7134感染P5-Rep78转基因Vero细胞系,结果表明Rep78的表达会抑制G207的增殖但是对ICP0突变的7134病毒没有明显的抑制作用.结果表明Rep不仅可以介导定点整合,同时也是AAV协助病毒的负调控物.更多还原

【Abstract】 AAV-2Rep78 is a multifunctional protein required for viral transcription,replication,and site-specific integration.It also mediates the interaction between AAV-2and its helper virus.We identified the cellular protein PML as a Rep78 interacting protein by co-immunoprecipitation.Further,based on the hybridization assay with an overlapped array of Rep protein and HEK293 cell lysate, the interacting fragment was located at545FPCRQCERM553 of Rep78specific C terminus.Transient expression of Rep78 resulted in an increased degradation of cellular PML,and the degradation could be partially recovered by treatment with MG132.Here,the HSV-1strain G207 containing ICP0and an ICP0-null mutant HSV-1were transfected in a P5-rep78 transgene Vero cell line,respectively,we observed an inhibitory effect of Rep78 expression on wide-type HSV-1replication,but not with the ICP0-null viral mutant HSV-1.The results suggested that Rep is not only a target,but also a contrary regulator for the helper virus of AAV.更多还原