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Molecular and Cellular Biology, July 2004, p. 6514-6524, Vol. 24, No. 14
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.14.6514-6524.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Mcm1 Promotes Replication Initiation by Binding Specific Elements at Replication Origins

Victoria K. Chang, Justin J. Donato, Clarence S. Chan, and Bik K. Tye^*

Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853

Received 15 February 2004/ Returned for modification 29 March 2004/ Accepted 19 April 2004

Minichromosome maintenance protein 1 (Mcm1) is required for efficient replication of autonomously replicating sequence (ARS)-containing plasmids in yeast cells. Reduced DNA binding activity in the Mcm1-1 mutant protein (P97L) results in selective initiation of a subset of replication origins and causes instability of ARS-containing plasmids. This plasmid instability in the mcm1-1 mutant can be overcome for a subset of ARSs by the inclusion of flanking sequences. Previous work showed that Mcm1 binds sequences flanking the minimal functional domains of ARSs. Here, we dissected two conserved telomeric X ARSs, ARS120 (XARS6L) and ARS131a (XARS7R), that replicate with different efficiencies in the mcm1-1 mutant. We found that additional Mcm1 binding sites in the C domain of ARS120 that are missing in ARS131a are responsible for efficient replication of ARS120 in the mcm1-1 mutant. Mutating a conserved Mcm1 binding site in the C domain diminished replication efficiency in ARS120 in wild-type cells, and increasing the number of Mcm1 binding sites stimulated replication efficiency. Our results suggest that threshold occupancy of Mcm1 in the C domain of telomeric ARSs is required for efficient initiation. We propose that origin usage in Saccharomyces cerevisiae may be regulated by the occupancy of Mcm1 at replication origins.

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* Corresponding author. Mailing address: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853. Phone: (607) 255-2445. Fax: (607) 255-2428. E-mail: bt16{at}cornell.edu.

Present address: Department of Physical Sciences, Kutztown University, Kutztown, PA 19530.

Present address: Section of Molecular Genetics and Microbiology, University of Texas, Austin, TX 78712-0162.

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Molecular and Cellular Biology, July 2004, p. 6514-6524, Vol. 24, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.14.6514-6524.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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