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Science 1 March 1996:
Vol. 271. no. 5253, pp. 1270 - 1272
DOI: 10.1126/science.271.5253.1270
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Reports

A Distinct G1 Step Required to Specify the Chinese Hamster DHFR Replication Origin

Jia-Rui Wu and David M. Gilbert *

Nuclei isolated from Chinese hamster ovary (CHO) cells at various times during the G1 phase of the cell cycle were stimulated to enter S phase by incubation in Xenopus egg cytosol. Replication of DNA initiated within the dihydrofolate reductase (DHFR) origin locus in nuclei isolated late in G1, but at random sites in nuclei isolated early in G1. A discrete transition point occurred 3 to 4 hours after metaphase. Neither replication licensing nor nuclear assembly was sufficient for origin recognition. Thus, a distinct cell cycle-regulated event in the nucleus restricts the initiation of replication to specific sites downstream of the DHFR gene.

Department of Biochemistry and Molecular Biology, SUNY Health Science Center, 750 East Adams Street, Syracuse, NY 13210, USA.
* To whom correspondence should be addressed.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
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Excess Mcm2-7 license dormant origins of replication that can be used under conditions of replicative stress.
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The promoter of the Chinese hamster ovary dihydrofolate reductase gene regulates the activity of the local origin and helps define its boundaries.
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Mammalian nuclei become licensed for DNA replication during late telophase.
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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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Activation of dormant origins of DNA replication in budding yeast.
C. Santocanale, K. Sharma, and J. F.X. Diffley (1999)
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DNA Methylation at Mammalian Replication Origins.
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J. Biol. Chem. 274, 25792-25800
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Mcm2, but Not RPA, Is a Component of the Mammalian Early G1-Phase Prereplication Complex.
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J. Cell Biol. 146, 709-722
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Nucleoskeleton and initiation of DNA replication in metazoan cells.
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J. Cell Sci. 111, 3663-3673
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E3-Ubiquitin Ligase/E6-AP Links Multicopy Maintenance Protein 7 to the Ubiquitination Pathway by a Novel Motif, the L2G Box.
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Identification of Primary Initiation Sites for DNA Replication in the Hamster Dihydrofolate Reductase Gene Initiation Zone.
T. Kobayashi, T. Rein, and M. L. DePamphilis (1998)
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Regulation of mammalian replication origin usage in Xenopus egg extract.
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ORC-dependent and origin-specific initiation of DNA replication at defined foci in isolated yeast nuclei..
P Pasero, D Braguglia, and S M Gasser (1997)
Genes & Dev. 11, 1504-1518
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Absence of an Unusual "Densely Methylated Island" at the Hamster dhfr ori-beta.
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Cell Cycle Control of DNA Replication.
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