Subclasses of simian virus 40 large T antigen: differential binding of two subclasses of T antigen from productively infected cells to viral and cellular DNA.
Two major subclasses of simian virus 40 (SV40) large T antigen were separated by zone velocity sedimentation of crude extracts from productively infected cells. These subclasses, which have been shown to differ biologically and biochemically ( Fanning et al., 1981), sedimented at 5-6S and 14-16S. The amount of T antigen in each form was estimated by complement fixation and by immunoprecipitation of T antigen from extracts of cells chronically labeled with [35S]methionine. Each form of T antigen was tested for specific binding to end-labeled restriction fragments of SV40 DNA using an immunoprecipitation assay. The 5-6S and 14-16S forms of T antigen both bound specifically to DNA sequences in the SV40 HindIII C fragment. The sequences required for binding both forms were localized in the same 35-bp region of the origin. However, significant differences in binding activity and affinity for specific and nonspecific DNA were demonstrated. These properties suggest that T antigen subclasses may serve different functions in the lytically infected cell.
D000818:Animals; D000952:Antigens, Polyomavirus Transforming; D002460:Cell Line; D004247:DNA / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D011485:Protein Binding; D011494:Protein Kinases / Q000235:genetics*; D013539:Simian virus 40 / Q000201:enzymology / Q000235:genetics*; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D014764:Viral Proteins; D004247:DNA; D011494:Protein Kinases