Chromatin Association of Human Origin Recognition Complex, Cdc6, and Minichromosome Maintenance Proteins during the Cell Cycle: Assembly of Prereplication Complexes in Late Mitosis

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Méndez, Juan, Stillman, Bruce·2000·Molecular and Cellular Biology

DOI 10.1128/mcb.20.22.8602-8612.2000

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Abstract

Evidence obtained from studies with yeast and Xenopus indicate that the initiation of DNA replication is a multistep process. The origin recognition complex (ORC), Cdc6p, and minichromosome maintenance (MCM) proteins are required for establishing prereplication complexes, upon which initiation is triggered by the activation of cyclin-dependent kinases and the Dbf4p-dependent kinase Cdc7p. The identification of human homologues of these replication proteins allows investigation of S-phase regulation in mammalian cells. Using centrifugal elutriation of several human cell lines, we demonstrate that whereas human Orc2 (hOrc2p) and hMcm proteins are present throughout the cell cycle, hCdc6p levels vary, being very low in early G(1) and accumulating until cells enter mitosis. hCdc6p can be polyubiquitinated in vivo, and it is stabilized by proteasome inhibitors. Similar to the case for hOrc2p, a significant fraction of hCdc6p is present on chromatin throughout the cell cycle, whereas hMcm proteins alternate between soluble and chromatin-bound forms. Loading of hMcm proteins onto chromatin occurs in late mitosis concomitant with the destruction of cyclin B, indicating that the mitotic kinase activity inhibits prereplication complex formation in human cells.

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Authors: Méndez, J., & Stillman, B.
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 2000-11-01
Volume: 20
Issue: 22
Pages: 8602-8612
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.20.22.8602-8612.2000
OpenAlex: W2095934571
Cited by: 915
References: 0
FWCI: 8.56
Type: journal article
Language: en
Open access: green
Licence: journals.asm.org/non-commercial-tdm-license (tdm)

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Chromatin Association of Human Origin Recognition Complex, Cdc6, and Minichromosome Maintenance Proteins during the Cell Cycle: Assembly of Prereplication Complexes in Late Mitosis

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