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Molecular and Cellular Biology, February 2005, p. 1560-1568, Vol. 25, No. 4
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.4.1560-1568.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Involvement of Human MCM8 in Prereplication Complex Assembly by Recruiting hcdc6 to Chromatin

Melanie Volkening1 and Ingrid Hoffmann1*

Cell Cycle Control and Carcinogenesis, DKFZ, German Cancer Research Center, Heidelberg, Germany1

Received 22 October 2004/ Accepted 10 November 2004

The MCM2-MCM7 complex is an essential component of the prereplication complex (pre-RC), which is recruited by the cdc6 and cdt1 proteins to origins of DNA replication during G1 phase. Here, we report that the accumulation on chromatin of another member of the MCM protein family, human MCM8 (hMCM8), occurs during early G1 phase, before the hMCM2-hMCM7 complex binds. hMCM8 interacts in vivo with two components of the pre-RC, namely, hcdc6 and hORC2. Depletion of endogenous hMCM8 protein by RNA interference leads to a delay of entry into S phase, suggesting a role for hMCM8 in G1 progression. Furthermore, down-regulation of hMCM8 also leads to a reduced loading of hcdc6 and the hMCM2-hMCM7 complex on chromatin. These results suggest that hMCM8 is a crucial component of the pre-RC and that the interaction between hMCM8 and hcdc6 is required for pre-RC assembly.

* Corresponding author. Mailing address: Cell Cycle Control and Carcinogenesis, F045 DKFZ, German Cancer Research Center, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany. Phone: 49-6221-424909. Fax: 49-6221-424902. E-mail: I.Hoffmann{at}dkfz.de.

Molecular and Cellular Biology, February 2005, p. 1560-1568, Vol. 25, No. 4
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.4.1560-1568.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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