Purification, properties, and subcellular localization of foxtail mosaic potexvirus 26-kDa protein.
The open reading frame 2 (ORF2) of the potexviral genome encodes a 24- to 26-kDa protein which is part of the "triple gene block," a group of overlapping ORFs also present in the genomes of the carla-, hordei-, and furoviruses. The product of these ORFs is believed to play a role in the cell-to-cell movement of the viruses in host plants. The amino acid sequences of the homologous ORF2 products encoded by these related viruses suggest that they specify NTP binding and possibly helicase activities. We have used an Escherichia coli expression system to produce significant amounts of the 26-kDa protein (p26) encoded by foxtail mosaic potexvirus ORF2. p28 was purified to near homogeneity by conventional purification methods and some of its biochemical properties were determined. We present evidence that p26 is an ATP, CTP, and RNA binding protein with apparent ATPase activity. Western blot analysis of infected plant extracts using a polyclonal antiserum produced against p26 indicates that it is a relatively stable protein maintained at high levels for at least 6 days following its peak level of expression. Moreover, it is found predominantly in the soluble fraction of infected tissues. An immunocytochemical analysis of infected Chenopodium quinoa leaves reveals that p26 is exclusively associated with cytoplasmic inclusions in proximity to but distinct from aggregates of viral particles.
Rouleau
M
M
Department of Biochemistry, University of Western Ontario, London, Canada.
Smith
R J
RJ
Bancroft
J B
JB
Mackie
G A
GA
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000914:Antibodies, Viral; D000918:Antibody Specificity; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007181:Inclusion Bodies, Viral / Q000737:chemistry / Q000648:ultrastructure; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010944:Plants / Q000382:microbiology; D017863:Potexvirus / Q000737:chemistry*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012265:Ribonucleotides / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D012265:Ribonucleotides; D014764:Viral Proteins; D012313:RNA; D000251:Adenosine Triphosphatases