ORF85 of HearNPV encodes the per os infectivity factor 4 (PIF4) and is essential for the formation of the PIF complex

ORF85 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) encodes a homologue of the per os infectivity factor 4 (PIF-4) of Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV). In this paper, the functions of HA85, particularly in relation to oral infection and interactions with other PIFs were investigated. An ha85-disrupted recombinant HearNPV was generated and resulted in a complete loss of oral infectivity. Western blotting and co-immunoprecipitation (Co-IP) analyses suggested PIF1, PIF2, and PIF3 assemble into a PIF complex in HearNPV ODV. Although Western blotting and Co-IP did not show that HA85 is associated with the PIF complex, further analysis revealed the inactivation of ha85 led to the disruption of PIF complex. Yeast two hybridization analyses revealed that HA85 interacts with P74, PIF1, PIF2 and PIF3. In conclusion, HA85 is identified as the PIF4 of HearNPV and is proposed to participate in the formation of HearNPV PIF complex via associations with other PIFs.