Abstract
Objective: To investigate the growth-inhibitory effect of Rep proteins on cells. Methods: We generated Rep-expressing plasmids that containing wild-type rep genes or mutant rep genes with point mutations in their ATP-binding sites. We obtained several HeLa cell clones expressing Rep proteins constitutively by the regulator plasmid. The expression of each Rep protein was detected by Western blotting with an anti-Rep monoclonal antibody. Results: Clones that expressed Rep52 and Rep40 grew more slowly than those which exhibited no detectable expression of Rep or mutant Rep protein. In contrast, clone w-33, which proliferated as quickly as control HeLa cells, expressed only Rep40. By comparing the expression levels of Rep proteins in these clones, we found that the ratio of Rep40 to Rep52 gradually increased in the faster growing clones. Conclusion: These results suggest that while Rep52 expression retards cell proliferation, Rep40 promotes recovery from Rep52-induced HeLa cell growth inhibition.
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