Disaggregation and reconstitution of oligomeric complexes of simian virus 40 large T-antigen.

Biochemical properties of multiple species of simian virus 40 (SV40) large T-antigen produced in SV40-infected monkey cells were investigated by zonal sedimentation centrifugation of radiolabelled cell extracts on sucrose gradients. Two major subpopulations of T-antigen detected by immunoprecipitation and gel electrophoresis could be distinctly separated: low molecular weight forms ranging approximately between 5S and 10S and higher oligomeric forms at about 16S to 23S. Removal of divalent cations by chelating agents such as EDTA disassembled higher oligomers into low molecular weight forms (5S to 10S). Adding divalent cations in excess of the EDTA concentration reassembled the higher oligomeric forms, showing a sedimentation behaviour like T-antigen in untreated cell extracts. Our results are compatible with the hypothesis that divalent cation binding properties of T-antigen participate in the natural pathway of assembling multiple oligomeric species.