Interaction of Heliothis armigera nuclear polyhedrosis viral capsid protein with its host actin.
In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.
Lu
Song-Ya
SY
Institute of Virology, Wuhan University, Wuhan 430072, P.R. China.
Qi
Yi-Peng
YP
Ge
Guo-Qiong
GQ
D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D036022:Capsid Proteins; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D049452:Green Fluorescent Proteins; D007313:Insecta; D007915:Lepidoptera / Q000821:virology*; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013997:Time Factors; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014763:Viral Matrix Proteins / Q000378:metabolism
D000199:Actins; D036022:Capsid Proteins; D018076:DNA, Complementary; D008164:Luminescent Proteins; D014763:Viral Matrix Proteins; D049452:Green Fluorescent Proteins; C074549:p39 protein, nuclear polyhedrosis virus