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We have studied structural changes in the simian virus 40 (SV40) replication origin induced by SV40 large tumor antigen (T antigen). T-antigen-induced changes in origin DNA conformation can be visualized as specific and discrete topologic changes in origin DNA minicircles. We discovered three origin-T-antigen complexes defined by changes in DNA linking number. These complexes probably reflected essential early steps in the initiation of DNA replication since their formation required DNA sequences that are necessary for DNA replication but do not affect T-antigen binding. There are striking parallels between the T antigen-origin interactions uncovered by this assay and the interactions between the DnaA, -B, and -C proteins and the Escherichia coli replication origin, suggesting a significant evolutionary conservation in the mechanisms that initiate DNA replication.

Simian virus 40 (SV40) large tumor antigen causes stepwise changes in SV40 origin structure during initiation of DNA replication.