A novel zinc finger is required for Mcm10 homocomplex assembly.
Mcm10 is a DNA replication factor that interacts with multiple subunits of the MCM2-7 hexameric complex. We report here that Mcm10 self-interacts and assembles into large homocomplexes (approximately 800 kDa). A conserved domain of 210 amino acid residues is sufficient for mediating self-interaction and complex assembly. A novel zinc finger within the conserved domain, CX10CX11CX2H, is essential for the homocomplex formation. Mutant alleles with amino acid substitutions at conserved cysteines and histidine in the zinc finger fail to assemble homocomplexes. A defect in homocomplex assembly correlates with defects in DNA replication and cell growth in the mutants. These observations suggest that homocomplex assembly is essential for Mcm10 function. Multisubunit Mcm10 homocomplexes may provide the structural basis for Mcm10 to interact with multiple subunits of the MCM2-7 hexamer.
Cook
Craig R
CR
Department of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Kung
Guosheng
G
Peterson
Francis C
FC
Volkman
Brian F
BF
Lei
Ming
M
D000483:Alleles; D020816:Amino Acid Motifs; D000596:Amino Acids / Q000737:chemistry; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D002455:Cell Division; D002850:Chromatography, Gel; D002868:Chromosomal Proteins, Non-Histone; D003545:Cysteine / Q000737:chemistry; D004247:DNA / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D064110:Minichromosome Maintenance Proteins; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers; D001616:beta-Galactosidase / Q000378:metabolism
D000596:Amino Acids; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D005656:Fungal Proteins; C076365:MCM10 protein, S cerevisiae; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; D004247:DNA; D005982:Glutathione Transferase; D001616:beta-Galactosidase; C069199:MCM2 protein, S cerevisiae; D064110:Minichromosome Maintenance Proteins; D015032:Zinc; D003545:Cysteine