ORF390 of white spot syndrome virus genome is identified as a novel anti-apoptosis gene.
Apoptosis serves as an important defense strategy employed by host cells against viral invasion. Many viruses contain the anti-apoptotic genes to block the defense-by-death response of host cells. In this study, we tried to identify the putative anti-apoptotic genes in white spot syndrome virus (WSSV) genome. We confirmed that actinomycin D could induce apoptosis of shrimp primary cells. However, the apoptosis triggered by actinomycin D was inhibited by WSSV infection. As mutants of Autographa californica nucleopolyhedrovirus (AcMNPV), AcMNPVDelta35k/pol+ lacks a functional P35 gene undergoing apoptosis and its infection could induce Sf9 cell apoptosis. To identify the putative apoptotic suppressor gene of WSSV, overlapping cosmid clones representing the entire WSSV genome were individually cotransfected along with genome DNA of AcMNPVDeltaP35k/pol+. Using this marker rescue assay, a WSSV DNA fragment that was able to rescue AcMNPVDeltaP35k/pol+ infection in Sf9 cells was isolated. By further sequence analysis and rescue assay, the ORF390 was identified as a novel anti-apoptotic gene. The ORF displays two putative caspase9 cleavage sites LLVETDGPS, VKLEHDGSK, and a caspase3 cleavage site EEDEVDGVP. The ORF was cloned into the pIE1 vector and then the recombinant vector was transfected into Sf9 cells. The Sf9 cells did not show obvious characteristics of apoptosis when infected with AcMNPVDeltaP35k/pol+. And the transient expression of ORF390 allowed AcMNPVDeltaP35k/pol+ replication in Sf9 cells and resulted in the formation of polyhedra successfully. The results indicate that function of ORF390 in WSSV is a kind of apoptotic suppressor like P35 in AcMNPV.
Wang
Zhimin
Z
Key laboratory of Virology, Ministry of Education, College of Life Science, Wuhan University, Wuhan 430072, Hubei Province, PR China.
Hu
Longbo
L
Yi
Guohua
G
Xu
Hua
H
Qi
Yipeng
Y
Yao
Lunuang
L
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D003609:Dactinomycin / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral / Q000187:drug effects / Q000502:physiology*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000821:virology*; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D046848:White spot syndrome virus 1 / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*
C495144:ORF390 protein, shrimp white spot syndrome virus; D014764:Viral Proteins; D003609:Dactinomycin