Function analysis of Ac-PCNA and Sf-PCNA during the Autographa californica multiple nucleopolyhedrovirus infection process.

Abstract

The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) possesses a gene, ac-pcna or ac49, which encodes a protein with similarity to proliferating cell nuclear antigen (PCNA). Homologs of this gene code for DNA polymerase processivity factors and are essential in the DNA replication systems. But the function of ac-pcna still remains unclear. To define the function of Ac-pcna in AcMNPV and Sf-pcna in host Sf9 cells, Bac-to-Bac baculovirus expression system was used to generate two recombinant baculoviruses: AcMNPV-Ac-pcna-EGFP and AcMNPV-Sf-pcna-EGFP. Results indicated that AcMNPV-mediated overexpression of Ac-PCNA and Sf-PCNA could stimulate replication of AcMNPV genome in the host Sf9 cells. Meanwhile, either AcMNPV-Ac-pcna-EGFP or AcMNPV-Sf-pcna-EGFP had a significant stimulating effect on Sf9 genome replication during infection. We also found that Ac-PCNA and Sf-PCNA could promote the production of budded virus. Ac-PCNA could improve the transcription level of ie2 gene dramatically and further improved the transcription of late gene, for example 38K and vp39, at 12h p.i.. Moreover, insecticidal potency test showed that the larvae of Beet armyworm in the AcMNPV-Ac-pcna-EGFP and AcMNPV-Sf-pcna-EGFP groups had a higher mortality rate (83.33 and 91.67%), a lower pupation rate (16.67 and 8.33%), and a lower emergence rate (6.67 and 3.33%), compared with those in AcMNPV-EGFP group. The function of Ac-PCNA and Sf-PCNA was confirmed in this study, which provided the theoretical foundation for using and modifying AcMNPV.