Salt-stable binding of the large T protein to DNA in polyoma virus chromatin.
Nucleoprotein complexes extracted from the nuclei of mouse cells lytically infected with polyoma virus contain an ATPase activity which appears to correspond to that of the viral large T protein, as it exhibits the same characteristic properties; in particular, the activity is extensively inhibited by polyclonal antibodies from animals bearing polyoma tumors (anti-T antigen antibodies) and by monoclonal antibodies against large T. Significant amounts of DNA were immunoprecipitated by adding these antibodies to the nucleoprotein complex, suggesting that the protein is tightly bound to DNA in the viral chromatin. Since one of the monoclonal antibodies quantitatively immunoprecipitated the pulse-labeled replicative intermediates, we conclude that some large T protein remains physically associated with the DNA throughout its replication cycle. After exposure to salt concentrations higher than 1 M KCl, about half of the large T-specific ATPase activity was still observed to co-sediment with 21S form I viral DNA. The observations that the sedimentation coefficient of the salt-stable complexes was shifted to 16S after a limited endonucleolytic digestion, and that both the viral DNA and the ATPase activity were co-precipitated in the presence of polyethylene glycol at high ionic strength, further demonstrated that the protein is engaged in an unusually stable complex with DNA in the viral chromatin.
D000251:Adenosine Triphosphatases / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000276:immunology; D000952:Antigens, Polyomavirus Transforming; D002843:Chromatin / Q000032:analysis; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism*; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000952:Antigens, Polyomavirus Transforming; D002843:Chromatin; D004279:DNA, Viral; D014764:Viral Proteins; D000251:Adenosine Triphosphatases