Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins Journal Article
| Authors: | Im, J. S.; Ki, S. H.; Farina, A.; Jung, D. S.; Hurwitz, J.; Lee, J. K. |
| Article Title: | Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins |
| Abstract: | In eukaryotes, the activation of the prereplicative complex and assembly of an active DNA unwinding complex are critical but poorly understood steps required for the initiation of DNA replication. In this report, we have used bimolecular fluorescence complementation assays in HeLa cells to examine the interactions between Cdc45, Mcm2-7, and the GINS complex (collectively called the CMG complex), which seem to play a key role in the formation and progression of replication forks. Interactions between the CMG components were observed only after the G1/S transition of the cell cycle and were abolished by treatment of cells with either a CDK inhibitor or siRNA against the Cdc7 kinase. Stable association of CMG required all three components of the CMG complex as well as RecQL4, Ctf4/And-1, and Mcm10. Surprisingly, depletion of TopBP1, a homologue of Dpb11 that plays an essential role in the chromatin loading of Cdc45 and GINS in yeast cells, did not significantly affect CMG complex formation. These results suggest that the proteins involved in the assembly of initiation complexes in human cells may differ somewhat from those in yeast systems. |
| Keywords: | unclassified drug; human cell; dna-binding proteins; nonhuman; cell cycle proteins; chromosomal proteins, non-histone; cell cycle; cell cycle s phase; complex formation; protein assembly; protein depletion; fluorescence; protein; small interfering rna; protein interaction; hela cell; hela cells; assay; luminescent proteins; nuclear proteins; bacterial proteins; recombinant fusion proteins; eukaryota; chromatin; protein-serine-threonine kinases; carrier proteins; cell therapy; multiprotein complexes; yeast; recq helicases; cyclin-dependent kinases; cyclin dependent kinase; phosphotransferase; cell cycle g1 phase; bimolecular fluorescence complementation; cdc45-mcm-gins complex; cdc7-dbf4 kinase; cyclin-dependent kinase; and 1 protein; cdc45 protein; cdc7 kinase; ctf4 protein; dpb11 protein; gins protein; mcm10 protein; mcm[2-7] protein; protein derivative; recql4 protein; topbp1 protein; yeast cell |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 106 |
| Issue: | 37 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2009-09-15 |
| Start Page: | 15628 |
| End Page: | 15632 |
| Language: | English |
| DOI: | 10.1073/pnas.0908039106 |
| PUBMED: | 19805216 |
| PROVIDER: | scopus |
| PMCID: | PMC2747170 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 13" - "Export Date: 30 November 2010" - "CODEN: PNASA" - "Source: Scopus" |
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