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Related Experiment Videos

Mcm1 promotes replication initiation by binding specific elements at replication origins.

Victoria K Chang1, Justin J Donato, Clarence S Chan

  • 1Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.

Molecular and Cellular Biology
|July 1, 2004
PubMed
Summary
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Minichromosome maintenance protein 1 (Mcm1) binding at replication origins is crucial for plasmid stability. Increased Mcm1 binding sites enhance replication efficiency, suggesting a threshold occupancy model for origin usage.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • DNA Replication

Background:

  • Minichromosome maintenance protein 1 (Mcm1) is essential for DNA replication initiation from autonomously replicating sequences (ARS) in yeast.
  • Mutations in Mcm1, like Mcm1-1 (P97L), reduce DNA binding, leading to selective origin firing and plasmid instability.
  • Flanking sequences can restore plasmid stability in Mcm1 mutants, indicating their role in Mcm1-mediated replication control.

Purpose of the Study:

  • To investigate the role of Mcm1 binding sites in the differential replication efficiencies of two telomeric X ARSs (ARS120 and ARS131a) in the mcm1-1 mutant.
  • To determine how Mcm1 occupancy at specific ARS elements influences replication initiation and plasmid stability.

Main Methods:

  • Comparative analysis of two telomeric ARS elements (ARS120 and ARS131a) in wild-type and mcm1-1 mutant yeast strains.

Related Experiment Videos

  • Site-directed mutagenesis to alter Mcm1 binding sites within the C domain of ARS120.
  • Assessment of plasmid replication efficiency and stability in response to Mcm1 binding site modifications.

    • Main Results:

    • ARS120, unlike ARS131a, showed efficient replication in the mcm1-1 mutant due to additional Mcm1 binding sites in its C domain.
    • Mutation of a key Mcm1 binding site in ARS120's C domain reduced its replication efficiency in wild-type cells.
    • Increasing the number of Mcm1 binding sites in ARS120 significantly enhanced its replication efficiency.

    Conclusions:

    • Threshold occupancy of Mcm1 in the C domain of telomeric ARSs is critical for efficient replication initiation.
    • Mcm1 binding site number and location play a regulatory role in Saccharomyces cerevisiae origin usage during DNA replication.