The cT mutation of PARA, an SV40-adenovirus 7 hybrid virus, abolishes the transport of SV40 T-antigen (T-ag) to the nucleus and results in the accumulation of T-ag in the cytoplasm of infected and transformed cells. The effect of the cT mutation on the association of T-ag with the cell surface was examined in PARA(cT)-infected monkey cells. Surface-associated T-ag was detected by immunofluorescence,lactoperoxidase-catalyzed cell-surface iodination, and cellular fractionation. Surface-associated T-ag was more readily labeled by cell-surface iodination with PARA(cT)-infected cells than with SV40 and wild-type PARA [PARA(nT)]-infected cells. Control experiments eliminated the possibility that the enhanced detection of T-ag in PARA(cT)-infected cells was due to the labeling of intracellular T-ag by lactoperoxidase or to T-ag leaking from dead cells and adhering to the cell surface. The elevated level of surface-associated T-ag in PARA(cT)-infected cells was confirmed by metabolic labeling and cellular fractionation, indicating that the increased iodination of surface-associated T-ag was due to a greater number of T-ag polypeptides on the surface rather than a greater exposure of tyrosine residues. Pulse-chase experiments demonstrated that the elevated level of surface-associated T-ag in PARA(cT)-infected cells was not due to a prolonged stability of surface-associated T-ag. The cT mutation provides a useful tool for studies of mechanisms governing intracellular protein transport.
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