Tsakraklides V and Bell SP (2010)

Abstract

The pre-replicative complex (pre-RC) is formed at all potential origins of replication through the action of the origin recognition complex (ORC), Cdc6, Cdt1, and the Mcm2-7 complex. The end result of pre-RC formation is the loading of the Mcm2-7 replicative helicase onto origin DNA. We examined pre-RC formation in vitro and found that it proceeds through separable binding events. Origin-bound ORC recruits Cdc6, and this ternary complex then promotes helicase loading in the presence of a pre-formed Mcm2-7-Cdt1 complex. Using a stepwise pre-RC assembly assay, we investigated the fate of pre-RC components during later stages of the reaction. We determined that helicase loading is accompanied by dissociation of ORC, Cdc6, and Cdt1 from origin DNA. This dissociation requires ATP hydrolysis at a late stage of pre-RC assembly. Our results indicate that pre-RC formation is a dynamic process.

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Reference Type: Journal Article | Research Support, N.I.H., Extramural | Research Support, Non-U.S. Gov't
Authors: Tsakraklides V, Bell SP
Primary Lit For: CDC6 | MCM2 | MCM6 | TAH11 | ORC2 | MCM3 | ORC6 | MCM5 | ORC4 | ORC1 | MCM4 | ... Show all
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