Article
- The EMBO Journal (2006) 25, 2209 - 2218
- doi:10.1038/sj.emboj.7601093
Published online: 20 April 2006
Subject Categories:
Communication between subunits within an archaeal clamp-loader complex
Anja Seyberta, Martin R Singletona, Nicola Cook, David R Hall and Dale B Wigley
- Clare Hall Laboratories, Cancer Research UK, London Research Institute, South Mimms Potters Bar, Herts, UK
Correspondence to:
Dale B Wigley, Clare Hall Laboratories, Cancer Research UK, London Research Institute, Blanche Lane, South Mimms Potters Bar, Herts EN6 3LD, UK. Tel.: +44 207 269 3930; Fax: +44 207 269 3803; E-mail: Dale.Wigley@cancer.org.uk
aThese authors contributed equally to this work
Received 5 January 2006; Accepted 22 March 2006
Abstract
We have investigated the communication between subunits in replication factor C (RFC) from Archaeoglobus fulgidus. Mutation of the proposed arginine finger in the small subunits results in a complex that can still bind ATP but has impaired clamp-loading activity, a process that normally only requires binding of nucleotide. The small subunit alone forms a hexameric ring that is six-fold symmetric in the absence of ATP. However, this symmetry is broken when the nucleotide is bound to the complex. A conformational change associated with nucleotide binding may relate to the opening of PCNA rings by RFC during the loading reaction. The structures also reveal the importance of the N-terminal helix of each subunit at the ATP-binding site. Analysis of mutant protein complexes containing subunits lacking this N-terminal helix reveals key distinct regulatory roles during clamp loading that are different for the large and small subunits in the RFC complex.
Keywords:
- clamp-loader,
- DNA replication,
- PCNA,
- RFC
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