Identification of functional domains required for HearNPV P10 filament formation.
Baculovirus encoded P10 form fibrillar structures in infected cells. We have tried to identify the functional domains for the P10 filament formation by green fluorescence protein (GFP) tag. The p10 gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was the subject of these studies. Different lengths of HearNPV p10 gene were constructed with GFP fused in frame to the C-terminus. The constructs were transfected into insect and mammalian cells and observed by confocal microscopy. The results indicated that the first N-terminal 66 amino acids, which form the complete coiled-coil domain, were necessary for the aggregation and formation of filament structures of HearNPV P10. The proline-rich region and the C-terminal positively charged amino acids were not necessary for the formation of fibrillar structure but had some impact on the shape of the fibrillar structures. No other baculoviral proteins were needed for the formation of P10 filament structures in transfected cells.
Dong
Chunsheng
C
State Key Laboratory of Virology, Key Laboratory of Molecular Virology and Joint-lab of Invertebrate Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
Li
Dan
D
Long
Gang
G
Deng
Fei
F
Wang
Hualin
H
Hu
Zhihong
Z
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D018411:Spodoptera; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C078072:p10 protein, Baculovirus