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Sequence-independent DNA binding and replication initiation by the human origin recognition complex

By Sanjay Vashee, Christin Cvetic, Wenyan Lu, Pamela Simancek, Thomas J. Kelly and Johannes C. Walter

Abstract

We report that a highly purified human origin recognition complex (HsORC) has intrinsic DNA-binding activity, and that this activity is modestly stimulated by ATP. HsORC binds preferentially to synthetic AT-rich polydeoxynucleotides, but does not effectively discriminate between natural DNA fragments that contain known human origins and control fragments. The complex fully restores DNA replication to ORC-depleted Xenopus egg extracts, providing strong evidence for its initiator function. Strikingly, HsORC stimulates initiation from any DNA sequence, and it does not preferentially replicate DNA containing human origin sequences. These data provide a biochemical explanation for the observation that in metazoans, initiation of DNA replication often occurs in a seemingly random pattern, and they have important implications for the nature of human origins of DNA replication

Topics: Research Papers
Publisher: Cold Spring Harbor Laboratory Press
Year: 2003
DOI identifier: 10.1101/gad.1084203
OAI identifier: oai:pubmedcentral.nih.gov:196240
Provided by: PubMed Central
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