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A mutant that affects the function of autonomously replicating sequences in yeast.

P Sinha, V Chang, B K Tye

    Journal of Molecular Biology
    |December 20, 1986
    PubMed
    Summary

    The mcm2 mutation in yeast disrupts minichromosome stability by affecting DNA replication initiation at autonomously replicating sequences (ARS). This defect is temperature-dependent and leads to plasmid loss and increased recombination.

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    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • DNA Replication

    Background:

    • Minichromosomes are essential for studying DNA replication and stability in yeast.
    • Minichromosome maintenance (mcm) mutants were previously identified as defective in plasmid stability.

    Purpose of the Study:

    • To investigate the role of the mcm2 mutation in yeast minichromosome stability.
    • To determine the effect of the mcm2 mutation on autonomously replicating sequences (ARS) and chromosomal elements.

    Main Methods:

    • Phenotypic analysis of mcm2 mutant yeast strains carrying various minichromosomes and plasmids.
    • Assessment of plasmid stability, segregation, and mitotic recombination frequencies.
    • Temperature-shift experiments to evaluate the temperature-dependent nature of the mcm2 defect.

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    Main Results:

    • The mcm2 mutation destabilizes minichromosomes, affecting both circular and linear plasmids lacking centromeres.
    • The defect is temperature-dependent and impacts all tested autonomously replicating sequences (ARS) at high temperatures.
    • mcm2 mutation leads to minichromosome loss and increased mitotic recombination, suggesting a defect in replication initiation.

    Conclusions:

    • The mcm2 mutation impairs the initiation of DNA replication at ARS elements, which function as chromosomal replication origins.
    • This replication initiation defect underlies the observed minichromosome instability and increased recombination in mcm2 mutants.