Functional analysis of Spodoptera litura nucleopolyhedrovirus p49 gene during Autographa californica nucleopolyhedrovirus infection of SpLi-221 cells.
The Spodoptera litura nucleopolyhedrovirus (SpltNPV) antiapoptotic gene Splt-p49 is able to rescue replication of a p35-null Autographa californica nucleopolyhedrovirus (AcMNPV) in AcMNPV-permissive Sf9 cells. In this study, an AcMNPV p35 knockout bacmid was generated through ET homologous recombination in Escherichia coli and designated as vAc(P35-KO). The Splt-p49 gene was transposed into the polyhedrin locus of vAc(P35-KO) to investigate if Splt-p49 has any antiapoptotic activity in the context of AcMNPV infection of AcMNPV-nonpermissive SpLi-221 cells. Our results demonstrated that Splt-p49 could not inhibit the apoptosis induced by AcMNPV infection of SpLi-221 cells when it was under the control of its native promoter. Western blot analysis showed that Splt-P49 was poorly expressed. When the expression of Splt-P49 was under the control of Drosophila hsp70 promoter, the expression of Splt-P49 was advanced, and a higher level of Splt-P49 was detected. As a result, the apoptosis of SpLi-221 cells was inhibited; however, budded-virus production did not improve in comparison with that in AcMNPV-infected SpLi-221 cells. These data indicated that there are other barrier(s) to AcMNPV replication in the nonpermissive SpLi-221 cells besides apoptosis.
Lin
Tiehao
T
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.
Yu
Mei
M
Wu
Wenbi
W
Yu
Qian
Q
Weng
Qingbei
Q
Yang
Kai
K
Yuan
Meijin
M
Pang
Yi
Y
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology*; D011401:Promoter Regions, Genetic; D018411:Spodoptera / Q000166:cytology / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D014764:Viral Proteins