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TheAutographa californica M nucleopolyhedrovirus (AcMNPV)orf79 (ac79 ) gene is a conserved gene in baculoviruses and shares homology with genes in ascoviruses, iridoviruses, and several bacteria. Ac79 has a conserved motif and structural similarities to UvrC and intron-encoded endonucleases. Ac79 is produced at early times during infection and concentrates in the nucleus of infected cells at late times, suggesting a cellular compartment-specific function. To investigate its function, anac79 -knockout bacmid was generated through homologous recombination inEscherichia coli . Titration assays showed that budded virus (BV) production was reduced in theac79 -knockout virus compared to control viruses, following either virus infection or the transfection of bacmid DNA. Theac79 -knockout virus-infected cells produced plaques smaller than those infected with controlac79 -carrying viruses. No obvious differences were observed in viral DNA synthesis, viral protein accumulation, or the formation of occlusion bodies inac79 -knockout and control viral DNA-transfected cells, indicating progression into the late and very late phases of viral infection. However, comparative analyses of the amounts of BV genomic DNA and structural proteins in a given quantity of infectious virions suggested that theac79 -knockout virus produced more noninfectious BV in infected cells than the control virus. The structure of theac79 -knockout BV determined by transmission electron microscopy appeared to be similar to that of the control virus, although aberrant capsid protein-containing tubular structures were observed in the nuclei ofac79 -knockout virus-infected cells. Tubular structures were not observed forac79 viruses with mutations in conserved endonuclease residues. These results indicate that Ac79 is required for efficient BV production.

journal of virology

The Autographa californica M Nucleopolyhedrovirus <i>ac79</i> Gene Encodes an Early Gene Product with Structural Similarities to UvrC and Intron-Encoded Endonucleases That Is Required for Efficient Budded Virus Production

The Autographa californica M Nucleopolyhedrovirus ac79 Gene Encodes an Early Gene Product with Structural Similarities to UvrC and Intron-Encoded Endonucleases That Is Required for Efficient Budded Virus Production