Involvement of transcriptional co-activator p300 in upregulated expression of HSP70 by aquareovirus non-structural protein NS31

Abstract

Members of Aquareovirus genus, including grass carp reovirus (GCRV), contribute to a serious threat to aquaculture animals accompanied by stress response. Our previous reports revealed that GCRV nonstructural protein NS31 serves as a potent contributor for virus selectively up-regulating specific heat shock protein 70-kd gene（HSP70），however，the mechanism by which inducing HSP70 gene expression is unknown. In this study, we further found that either the N- or C-terminal domain of GCRV NS31 is responsible for enhancing fish HSP70 promoter transcription, and recombinant NS31 protein purified from baculovirus expression system seems to not directly bind HSP70 basic promoter in vitro by an electrophoretic mobility shift assay. However, the transcriptional co-activator p300 was identified as a potential interacting partner for NS31 by pull-down assay. Moreover, knock-down of p300 or addition of p300 inhibitor resulted in obviously reduced HSP70 expression by NS31 or GCRV infection suggesting that the well-characterized heat-shock-responsive HSF1/p300 transcriptional complex might involve in the induction of HSP70. These results collectively reveal this aquareovirus generates cell stress response through its nonstructural protein NS31 recruiting transcriptional co-activator p300.