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>Sequential and ordered assembly of e1 initiator complexes on the papillomavirus origin of DNA replication generates progressive structural changes related to melting.
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Sequential and ordered assembly of e1 initiator complexes on the papillomavirus origin of DNA replication generates progressive structural changes related to melting.
Multiple binding sites for an initiator protein are a common feature of replicator sequences from various organisms. By binding to the replicator, initiators mark the site and contribute to melting or distortion of the DNA by largely unknown mechanisms. Here we analyze origin of DNA replication (ori) binding by the E1 initiator and show sequential binding to a set of overlapping binding sites. The assembly of these initiator complexes is controlled by a gradual reduction in the dependence of interactions between the initiator and DNA and a gradual increase in the reliance on interactions between initiator molecules, providing a mechanism for sequential and orderly assembly. Importantly, the binding of the initiator causes progressive structural alterations both in the sites and in the sequences flanking the sites, eventually generating severe structural alterations. These results indicate that the process of template melting may be incremental, where binding of each initiator molecule serves as a wedge that upon binding gradually alters the template structure. This mechanism may explain the requirement for multiple initiator binding sites that is observed in many ori's.
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Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724. Graduate Program in Genetics, State University of New York at Stony Brook, Stony Brook, New York 11794.;
