Multiple isoelectric forms of poliovirus RNA-dependent RNA polymerase: evidence for phosphorylation.
Poliovirus-specific RNA-dependent RNA polymerase (3Dpol) was purified to apparent homogeneity. A single polypeptide of an apparent molecular weight of 63,000 catalyzes the synthesis of dimeric and monomeric RNA products in response to the poliovirion RNA template. Analysis of purified 3Dpol by two-dimensional electrophoresis showed multiple forms of 3Dpol, suggesting posttranslational modification of the protein in virus-infected cells. The two major forms of 3Dpol appear to have approximate pI values of 7.1 and 7.4. Incubation of purified 3Dpol with calf intestinal phosphatase resulted in almost complete disappearance of the pI 7.1 form and a concomitant increase in the intensity of the pI 7.4 form of 3Dpol. Addition of 32P-labeled Pi during infection of HeLa cells with poliovirus resulted in specific labeling of 3Dpol and 3CD, a viral protein which contains the entire 3Dpol sequence. Both 3Dpol and 3CD appear to be phosphorylated at serine residues. Ribosomal salt washes prepared from both mock- and poliovirus-infected cells contain phosphatases capable of dephosphorylating quantitatively the phosphorylated form (pI 7.1) of 3Dpol.
Ransone
L J
LJ
Department of Microbiology and Immunology, University of California, School of Medicine, Los Angeles 90024-1747.
Dasgupta
A
A
D000596:Amino Acids / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D017955:Poliovirus / Q000201:enzymology*; D012330:RNA, Double-Stranded / Q000096:biosynthesis; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism
D000596:Amino Acids; D007527:Isoenzymes; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases