Structural analysis of a eukaryotic sliding DNA clamp-clamp loader complex

• Published in: Nature 2004-06, Vol.429 (6993), p.724-730
• Main Authors: Kuriyan, John, Bowman, Gregory D, O'Donnell, Mike
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - analogs & derivatives; Adenosine Triphosphate - metabolism; ATP; Binding Sites; Biological and medical sciences; Crystalline structure; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - metabolism; Eukaryotic Cells; Fundamental and applied biological sciences. Psychology; Humanities and Social Sciences; Models, Molecular; Molecular biology; Molecular biophysics; multidisciplinary; Nucleic Acid Conformation; Proliferating Cell Nuclear Antigen - chemistry; Proliferating Cell Nuclear Antigen - metabolism; Protein Conformation; Protein Subunits - chemistry; Protein Subunits - metabolism; Replication Protein C; Saccharomyces cerevisiae; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - metabolism; Science; Science (multidisciplinary); Structural analysis; Structure in molecular biology; Yeast; Yeasts
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/nature02585

Sliding clamps are ring-shaped proteins that encircle DNA and confer high processivity on DNA polymerases. Here we report the crystal structure of the five-protein clamp loader complex (replication factor-C, RFC) of the yeast Saccharomyces cerevisiae , bound to the sliding clamp (proliferating cell nuclear antigen, PCNA). Tight interfacial coordination of the ATP analogue ATP-γS by RFC results in a spiral arrangement of the ATPase domains of the clamp loader above the PCNA ring. Placement of a model for primed DNA within the central hole of PCNA reveals a striking correspondence between the RFC spiral and the grooves of the DNA double helix. This model, in which the clamp loader complex locks onto primed DNA in a screw-cap-like arrangement, provides a simple explanation for the process by which the engagement of primer–template junctions by the RFC:PCNA complex results in ATP hydrolysis and release of the sliding clamp on DNA.