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Structural basis for DNA binding by replication initiator Mcm10

Eric M Warren1, Sivaraja Vaithiyalingam, Justin Haworth

  • 1Department of Biological Sciences, Vanderbilt University, Nashville, TN 37232, USA.

Structure (London, England : 1993)
|December 17, 2008
PubMed

Insights

Mcm10, a key DNA replication protein, uses a unique zinc finger to bind single-stranded DNA. This interaction is crucial for replication and preventing stress in cells.

Area of Science:

  • Molecular Biology
  • Structural Biology
  • Biochemistry

Background:

  • Mcm10 is vital for eukaryotic DNA replication fork assembly and progression.
  • Mcm10's internal domain (Mcm10-ID) interacts with single-stranded DNA, DNA polymerase alpha, and PCNA.

Purpose of the Study:

  • To determine the crystal structure of Xenopus laevis Mcm10-ID.
  • To elucidate the mechanism of Mcm10's single-stranded DNA binding.
  • To investigate the functional significance of Mcm10 DNA binding in vivo.

Main Methods:

  • X-ray crystallography to determine the Mcm10-ID structure.
  • NMR chemical shift perturbation and in vitro mutagenesis to study DNA binding.
  • Analysis of Mcm10 mutations in Saccharomyces cerevisiae to assess replication stress sensitivity.

Main Results:

  • The crystal structure reveals Mcm10-ID possesses an oligonucleotide/oligosaccharide-fold followed by a basic zinc finger.
  • NMR and mutagenesis studies demonstrate this region directly binds single-stranded DNA.
  • Mutations affecting this DNA binding in yeast increase sensitivity to replication stress.

Conclusions:

  • The Mcm10 zinc finger is essential for direct single-stranded DNA binding.
  • This DNA binding function is critical for cellular replication fidelity and stress response.
  • The structure provides a framework for understanding Mcm10's role in coordinating replisome components.

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