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Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2006 Jul 05; Vol. 103 (27), pp. 10236-10241. Date of Electronic Publication: 2006 Jun 23. - Publication Year :
- 2006
-
Abstract
- The protein Cdc45 plays a critical but poorly understood role in the initiation and elongation stages of eukaryotic DNA replication. To study Cdc45's function in DNA replication, we purified Cdc45 protein from Drosophila embryo extracts by a combination of traditional and immunoaffinity chromatography steps and found that the protein exists in a stable, high-molecular-weight complex with the Mcm2-7 hexamer and the GINS tetramer. The purified Cdc45/Mcm2-7/GINS complex is associated with an active ATP-dependent DNA helicase function. RNA interference knock-down experiments targeting the GINS and Cdc45 components establish that the proteins are required for the S phase transition in Drosophila cells. The data suggest that this complex forms the core helicase machinery for eukaryotic DNA replication.
- Subjects :
- Animals
Calcium-Calmodulin-Dependent Protein Kinases genetics
Cell Cycle Proteins genetics
Chromatography, Affinity
DNA biosynthesis
DNA Helicases genetics
Drosophila Proteins genetics
Drosophila melanogaster enzymology
Drosophila melanogaster genetics
Drosophila melanogaster metabolism
Eukaryotic Cells enzymology
Peptides pharmacology
Protein Binding
S Phase
Calcium-Calmodulin-Dependent Protein Kinases isolation & purification
Calcium-Calmodulin-Dependent Protein Kinases metabolism
Cell Cycle Proteins isolation & purification
Cell Cycle Proteins metabolism
DNA Helicases isolation & purification
DNA Helicases metabolism
DNA Replication
Drosophila Proteins isolation & purification
Drosophila Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0027-8424
- Volume :
- 103
- Issue :
- 27
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 16798881
- Full Text :
- https://doi.org/10.1073/pnas.0602400103