Open AccessCloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of the central zinc‐binding domain of the human Mcm10 DNA‐replication factor
Author(s) -
Jung Nam Young,
Bae Won Jin,
Chang Jeong Ho,
Kim Young Chang,
Cho Yunje
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108011640
Subject(s) - dna replication , replication factor c , origin recognition complex , eukaryotic dna replication , control of chromosome duplication , pre replication complex , crystallography , proliferating cell nuclear antigen , dna , biology , replication protein a , dna binding domain , chemistry , microbiology and biotechnology , dna binding protein , transcription factor , genetics , gene
The initiation of eukaryotic DNA replication requires the tightly controlled assembly of a set of replication factors. Mcm10 is a highly conserved nuclear protein that plays a key role in the initiation and elongation processes of DNA replication by providing a physical link between the Mcm2–7 complex and DNA polymerases. The central domain, which contains the CCCH zinc‐binding motif, is most conserved within Mcm10 and binds to DNA and several proteins, including proliferative cell nuclear antigen. In this study, the central domain of human Mcm10 was crystallized using the hanging‐drop vapour‐diffusion method in the presence of PEG 3350. An X‐ray diffraction data set was collected to a resolution of 2.6 Å on a synchrotron beamline. The crystals formed belonged to space group R 3, with unit‐cell parameters a = b = 99.5, c = 133.0 Å. According to Matthews coefficient calculations, the crystals were predicted to contain six MCM10 central domain molecules in the asymmetric unit.