P64, a novel major virion DNA-binding protein potentially involved in condensing the Spodoptera frugiperda Ascovirus 1a genome.
We recently identified 21 structural proteins in the virion of Spodoptera frugiperda ascovirus 1a (SfAV1a), a virus with a large, double-stranded DNA genome of 157 kbp, which attacks species of the lepidopteran family Noctuidae. The two most abundant virion proteins were the major capsid protein and a novel protein (P64) of 64 kDa that contained two distinct domains not known previously to occur together. The amino-terminal half of P64 (residues 1 to 263) contained four repeats (a recently recognized motif with an unknown function) of a virus-specific two-cysteine adaptor. Adjoined to this, the carboxy-terminal half of P64 (residues 279 to 455) contained 14 copies of a highly basic, tandemly repeated motif rich in arginine and serine, having an 11- to 13-amino-acid consensus sequence, SPSQRRSTS(V/K)(A/S)RR, yielding a predicted isoelectric point of 12.2 for this protein. In the present study, we demonstrate by Southwestern analysis that SfAV1a P64 was the only virion structural protein that bound DNA. Additional electrophoretic mobility shift assays showed that P64 bound SfAV1a as well as non-SfAV1a DNA. Furthermore, we show through immunogold labeling of ultrathin sections that P64 is a component of virogenic stroma and appears to be progressively incorporated into the SfAV1a DNA core during virion assembly. As no other virion structural protein bound DNA and no basic DNA-binding proteins of lower mass are encoded by the SfAV1a genome or were identified by proteomic analysis, our results suggest that P64's function is to condense the large genome of this virus and assist in packaging this genome into its virion.
Tan
Yeping
Y
Department of Entomology, University of California-Riverside, Riverside, CA 92521, USA.
Spears
Tatsinda
T
Bideshi
Dennis K
DK
Johnson
Jeffrey J
JJ
Hice
Robert
R
Bigot
Yves
Y
Federici
Brian A
BA
D000595:Amino Acid Sequence; D000818:Animals; D029223:Ascoviridae / Q000502:physiology*; D026501:Blotting, Southwestern; D036022:Capsid Proteins / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D016679:Genome, Viral; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D018411:Spodoptera / Q000821:virology; D019065:Virus Assembly
D036022:Capsid Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins