Structural basis for DNA binding by replication initiator Mcm10.

Warren EM, Vaithiyalingam S, Haworth J, Greer B, Bielinsky AK, Chazin WJ, Eichman BF
Structure. 2008 16 (12): 1892-901

PMID: 19081065 · PMCID: PMC2636851 · DOI:10.1016/j.str.2008.10.005

Mcm10 is an essential eukaryotic DNA replication protein required for assembly and progression of the replication fork. The highly conserved internal domain (Mcm10-ID) has been shown to physically interact with single-stranded (ss) DNA, DNA polymerase alpha, and proliferating cell nuclear antigen (PCNA). The crystal structure of Xenopus laevis Mcm10-ID presented here reveals a DNA binding architecture composed of an oligonucleotide/oligosaccharide-fold followed in tandem by a variant and highly basic zinc finger. NMR chemical shift perturbation and mutational studies of DNA binding activity in vitro reveal how Mcm10 uses this unique surface to engage ssDNA. Corresponding mutations in Saccharomyces cerevisiae result in increased sensitivity to replication stress, demonstrating the functional importance of DNA binding by this region of Mcm10 to replication. In addition, mapping Mcm10 mutations known to disrupt PCNA, polymerase alpha, and DNA interactions onto the crystal structure provides insight into how Mcm10 might coordinate protein and DNA binding within the replisome.

MeSH Terms (23)

Amino Acid Motifs Amino Acid Sequence Animals Binding Sites Biophysical Phenomena DNA DNA-Binding Proteins DNA Mutational Analysis DNA Polymerase I DNA Replication Minichromosome Maintenance Proteins Models, Biological Models, Molecular Molecular Sequence Data Mutation Nuclear Magnetic Resonance, Biomolecular Proliferating Cell Nuclear Antigen Protein Binding Protein Structure, Tertiary Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Zinc Fingers

Connections (3)

This publication is referenced by other Labnodes entities:

Links