Choristoneura fumiferana multiple nucleopolyhedrovirus LEF-3–P143 complex can complement DNA replication and budded virus in an AcMNPV LEF-3–P143 double knockout bacmid

Abstract

Transient replication assays usingAutographa californicamultiple nucleopolyhedrovirus (AcMNPV) andChoristoneura fumiferanamultiple nucleopolyhedrovirus (CfMNPV) genes suggested that the interactions between P143, the viral helicase and LEF-3, a ssDNA-binding protein, may represent virus species specificity determinants. P143 and LEF-3 are essential for DNA replication in these assays and together with IE-1, the major immediate-early transcription factor, may be part of the viral replisome. In the current report, alef-3/p143double-knockout AcMNPV bacmid was constructed that was defective for viral DNA replication and late gene expression. When the homologouslef-3/p143CfMNPV genes were introduced into this double-knockout bacmid, DNA replication was restored but the level of replication was lower, budded virus production was delayed, and the yields were reduced from those in an AcMNPV-rescue bacmid. These results suggest that to maximize virus replication, baculovirus replisome assembly and function requires protein–protein interactions between P143 and LEF-3, and other viral proteins.