Simian virus 40 large T antigen (large T) in the early and the late phases of infection differs significantly in its sequence-specific DNA-binding and ATPase activities, indicating that different large-T populations participate in virus-specific events at various stages of the infectious cycle. To further characterize these large-T populations, we have analyzed nuclear subclasses of large T, isolated from their in vivo location, for their biochemical activities. We show that chromatin- and nuclear matrix-associated large-T molecules exhibit different simian virus 40 control region (ORI) DNA-binding and ATPase activities. The association of large T with a certain nuclear substructure, therefore, subcompartmentalizes large-T molecules exerting different biochemical activities. Nuclear subcompartmentalization thus may provide a higher-order level for the regulation of biochemical activities of large T in vivo
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