Identification of protein-protein interactions of the occlusion-derived virus-associated proteins of Helicoverpa armigera nucleopolyhedrovirus

State Key Laboratory of Virology and Joint Laboratory of Invertebrate Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China Correspondence Zhihong Hu huzh{at}wh.iov.cn The purpose of this study was to identify protein–protein interactions among the components of the occlusion-derived virus (ODV) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV), a group II alphabaculovirus in the family Baculoviridae . To achieve this, 39 selected genes of potential ODV structural proteins were cloned and expressed in the Gal4 yeast two-hybrid (Y2H) system. The direct-cross Y2H assays identified 22 interactions comprising 13 binary interactions [HA9–ODV-EC43, ODV-E56–38K, ODV-E56–PIF3, LEF3–helicase, LEF3–alkaline nuclease (AN), GP41–38K, GP41–HA90, 38K–PIF3, 38K–PIF2, VP80–HA100, ODV-E66–PIF3, ODV-E66–PIF2 and PIF3–PIF2] and nine self-associations (IE1, HA44, LEF3, HA66, GP41, CG30, 38K, PIF3 and P24). Five of these interactions – LEF3–helicase and LEF3–AN, and the self-associations of IE1, LEF3 and 38K – have been reported previously in Autographa californica multiple nucleopolyhedrovirus. As HA44 and HA100 were two newly identified ODV proteins of group II viruses, their interactions were further confirmed. The self-association of HA44 was verified with a His pull-down assay and the interaction of VP80–HA100 was confirmed by a co-immunoprecipitation assay. A summary of the protein–protein interactions of baculoviruses reported so far, comprising 68 interactions with 45 viral proteins and five host proteins, is presented, which will facilitate our understanding of the molecular mechanisms of baculovirus infection. Present address: Laboratory of Virology, Wageningen University, Binnenhaven 11, 6709 PD Wageningen, The Netherlands. A supplementary table detailing the proteins and primers used for the Y2H assay is available with the online version of this paper.