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Cdc6 ATPase activity disengages Cdc6 from the pre-replicative complex to promote DNA replication

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Title: Cdc6 ATPase activity disengages Cdc6 from the pre-replicative complex to promote DNA replication
Authors: Speck, C
Item Type: Journal Article
Abstract: © 2015, Chang et al.To initiate DNA replication, cells first load an MCM helicase double hexamer at origins in a reaction requiring ORC, Cdc6, and Cdt1, also called pre-replicative complex (pre-RC) assembly. The essential mechanistic role of Cdc6 ATP hydrolysis in this reaction is still incompletely understood. Here, we show that although Cdc6 ATP hydrolysis is essential to initiate DNA replication, it is not essential for MCM loading. Using purified proteins, an ATPase-defective Cdc6 mutant ‘Cdc6-E224Q’ promoted MCM loading on DNA. Cdc6-E224Q also promoted MCM binding at origins in vivo but cells remained blocked in G1-phase. If after loading MCM, Cdc6-E224Q was degraded, cells entered an apparently normal S-phase and replicated DNA, a phenotype seen with two additional Cdc6 ATPase-defective mutants. Cdc6 ATP hydrolysis is therefore required for Cdc6 disengagement from the pre-RC after helicase loading to advance subsequent steps in helicase activation in vivo.
Issue Date: 25-Aug-2015
Date of Acceptance: 22-Jul-2015
URI: http://hdl.handle.net/10044/1/25914
DOI: https://dx.doi.org/10.7554/eLife.05795.001
ISSN: 2050-084X
Publisher: eLife Sciences Publications
Journal / Book Title: eLife
Volume: 4
Copyright Statement: © 2015, Chang et al This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
Publication Status: Published
Article Number: e05795
Appears in Collections:Institute of Clinical Sciences

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