An ac34 deletion mutant of Autographa californica nucleopolyhedrovirus exhibits delayed late gene expression and a lack of virulence in vivo.
Ac34 and its homologs are highly conserved in all sequenced alphabaculoviruses. In this paper, we show that ac34 transcripts were detected from 6 to 48 h postinfection (p.i.) in Autographa californica nucleopolyhedrovirus (AcMNPV)-infected Sf9 cells. Ac34 localized to both the cytoplasm and the nuclei of infected cells but was not a viral structural protein. To determine the function of ac34 in the viral life cycle, an ac34 knockout AcMNPV (vAc34KO) was constructed. Compared with wild-type and repair viruses, vAc34KO exhibited an approximately 100-fold reduction in infectious virus production. Further investigations showed that the ac34 deletion did not affect the replication of viral DNA, polyhedron formation, or nucleocapsid assembly but delayed the expression of late genes, such as vp39, 38k, and p6.9. Bioassays revealed that vAc34KO was unable to establish a fatal infection in Trichoplusia ni larvae via per os inoculation. Few infectious progeny viruses were detected in the hemolymph of the infected larvae, indicating that the replication of vAc34KO was attenuated. These results suggest that Ac34 is an activator protein that promotes late gene expression and is essential for the pathogenicity of AcMNPV.
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.
Yi
Y
Cai
Zhao
Z
Long
Jianxiang
J
Qiu
Meijin
M
Yuan
Guanghong
G
Li
Kai
K
Yang
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D001681:Biological Assay; D017353:Gene Deletion; D015870:Gene Expression; D005786:Gene Expression Regulation; D006458:Hemolymph / Q000378:metabolism; D007700:Kinetics; D007814:Larva / Q000821:virology; D007915:Lepidoptera / Q000378:metabolism; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017386:Sequence Homology, Amino Acid; D014774:Virulence