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The Mcm467 complex of Saccharomyces cerevisiae is preferentially activated by autonomously replicating DNA sequences

Esther E Biswas-Fiss1, Sujata M Khopde, Subhasis B Biswas

  • 1Department of Molecular Biology, School of Osteopathic Medicine, Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey, Stratford, New Jersey 08084, USA. subhasis.biswas@umdnj.edu

Biochemistry
|February 23, 2005
PubMed

Insights

Single-stranded DNA (ssDNA) from yeast replication origins specifically stimulates Mcm467 helicase activity. Replication protein A also enhances this Mcm467 complex

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Yeast Genetics

Background:

  • The Mcm467 complex is a key component of the DNA replication machinery in Saccharomyces cerevisiae.
  • Understanding the regulation of helicase activity is crucial for comprehending DNA replication initiation and progression.

Purpose of the Study:

  • To investigate the role of single-stranded DNA (ssDNA) sequences in modulating the ATPase and DNA helicase activities of the Mcm467 complex.
  • To identify specific ssDNA sequences that act as potent stimulators of Mcm467 complex function.

Main Methods:

  • Biochemical assays measuring ATPase activity of the Mcm467 complex in the presence of various ssDNA oligonucleotides.
  • DNA helicase activity assays using ARS1-derived and synthetic ssDNA sequences.
  • Analysis of sequence-specific stimulation by ssDNA derived from Saccharomyces cerevisiae autonomously replicating sequence 1 (ARS1).

Main Results:

  • Mcm467 ATPase activity is modulated in a sequence-specific manner by ssDNA.
  • ssDNA sequences from ARS1, particularly those containing A and B1 motifs, are highly effective stimulators.
  • Yeast replication protein A also stimulates Mcm467 ATPase activity.

Conclusions:

  • Specific ssDNA sequences, notably those from ARS1, activate the Mcm467 complex, enhancing its ATPase and DNA helicase activities.
  • The findings highlight the importance of ssDNA sequence composition in regulating DNA helicase function during replication initiation in yeast.

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