Host factor cyclophilin B affects Orf virus replication by interacting with viral ORF058 protein

Abstract

Poxviruses have evolved multiple strategies to modulate host-derived factors to create an optimal environment for viral efficient replication. Our previous study indicated that cyclophilin B (CypB) is a critical factor for ORFV replication in MDBK cells. However, the precise molecular mechanism by which CypB facilitates ORFV replication remains less understood. In the present study, the function of CypB in ORFV replication is further evaluated. The overexpression of CypB was observed to facilitate ORFV replication in OFTu cells and HeLa cells, however, RNA interference (RNAi)-mediated reduction of endogenous CypB decreased the levels of ORFV replication. Coimmunoprecipitation experiments revealed that the CypB interacted with ORFV ORF058 protein, a late protein involved in virus entry. The interaction of host factor CypB and ORF058 protein was further confirmed by confocal microscopy analysis and GST-pull down. In addition, the 52-55 aa was identified as the critical binding sites for CypB on ORF058 protein by GST-pull down with OFTu cells overexpressing CypB and purified GST-tagged truncated ORF058. In conclusion, we demonstrate that CypB is a critical host factor for ORFV replication in vitro by interacting with ORF058 protein, providing new insights into ORFV pathogenesis.

Keywords: Host factor CypB; Interaction; ORF058 protein; ORFV.