Identification of a hydrophobic domain of HA2 essential to morphogenesis of Helicoverpa armigera nucleopolyhedrovirus.
The HA2 protein of the Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearNPV) is a WASP homology protein capable of nucleating branched actin filaments in the presence of the Arp2/3 complex in vitro. To determine the role of ha2 in the HearNPV life cycle, ha2 knockout and ha2 repair bacmids were constructed. Transfection and infection analysis demonstrated that the ha2 null bacmid was unable to produce infectious budded virus (BV), while the repair bacmid rescued the defect. In vitro analysis demonstrated that the WCA domain of HA2 accelerates Arp2/3-mediated actin assembly and is indispensable to the function of HA2. However, analysis of the repaired recombinant with a series of truncated ha2 mutants demonstrated that the WCA domain was essential but not enough to yield infectious virions, and a hydrophobic domain (H domain) consisting of amino acids (aa) 167 to 193 played a pivotal role in the production of BV. Subcellular localization analysis with enhanced green fluorescent protein fusions showed that the H domain functioned as a nuclear localization signal. In addition, deletion of the C terminus of the ha2 product, a phosphatidylinositol 4-kinase homolog, dramatically decreased the viral titer, while deletion of 128 aa from the N terminus did not affect HA2 function.
Wang
Qian
Q
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, People's Republic of China.
Wang
Yun
Y
Liang
Changyong
C
Song
Jianhua
J
Chen
Xinwen
X
D000818:Animals; D002460:Cell Line; D017353:Gene Deletion; D005816:Genetic Complementation Test; D007915:Lepidoptera; D046529:Microscopy, Electron, Transmission; D019913:Nuclear Localization Signals / Q000235:genetics; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D019065:Virus Assembly; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics / Q000502:physiology*
C526765:HA2 protein, Helicoverpa armigera nucleopolyhedrovirus; D019913:Nuclear Localization Signals; D014764:Viral Proteins; D051316:Wiskott-Aldrich Syndrome Protein