Biochemical characterization of the structural and nonstructural polypeptides of a porcine group C rotavirus.
Purified virions or radiolabeled lysates of infected MA104 cells were used to characterize the structural and nonstructural polypeptides of a porcine group C rotavirus. At least six structural proteins were identified from purified group C rotavirus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Of these, two (37,000- and 33,000-molecular-weight polypeptides) were associated with the outer shell, as demonstrated by the ability of EDTA to remove them from the purified virion. The other four polypeptides (molecular weights, 125,000, 93,000, 74,000, and 41,000) were located in the inner shell. The structural or nonstructural nature of a 25,000-molecular-weight protein identified in our studies was unclear. Glycosylation inhibition studies with tunicamycin in infected cells demonstrated that the 37,000- and 25,000-molecular-weight proteins were glycosylated and contained mannose-rich oligosaccharides identified by radiolabeling of the infected cells with [3H]mannose. The 37,000-molecular-weight outer shell glycoprotein was shown by pulse-chase experiments to be posttranslationally processed. The kinetics of viral polypeptide synthesis in infected cells were also studied, and maximal synthesis occurred at 6 to 9 h postinfection. The 41,000-molecular-weight inner capsid polypeptide was the most abundant and was the subunit structure of a 165,000-molecular-weight protein aggregate. Two polypeptides (molecular weights, 39,000 and 35,000) appeared to be nonstructural, as determined by comparison of the protein pattern of radiolabeled infected cell lysates with that of purified virions. Radioimmunoprecipitation was used to examine the serologic cross-reactions between the viral polypeptides of a group C rotavirus with those of a group A rotavirus. No serologic cross-reactivities were detected. The polypeptides of group A and C rotaviruses are compared and discussed.
Jiang
B M
BM
Food Animal Health Research Program, Ohio State University, Wooster 44691-4096.
Saif
L J
LJ
Kang
S Y
SY
Kim
J H
JH
D000818:Animals; D002213:Capsid / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D008970:Molecular Weight; D011233:Precipitin Tests; D012401:Rotavirus / Q000032:analysis* / Q000378:metabolism; D013552:Swine / Q000382:microbiology; D014758:Viral Core Proteins / Q000032:analysis; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000032:analysis* / Q000096:biosynthesis
D046911:Macromolecular Substances; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins