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- Title
The architecture of an Okazaki fragment-processing holoenzyme from the archaeon Sulfolobus solfataricus.
- Authors
Cannone, Giuseppe; Yuli Xu; Beattie, Thomas R.; Bell, Stephen D.; Spagnolo, Laura
- Abstract
DNA replication on the lagging strand occurs via the synthesis and maturation of Okazaki fragments. In archaea and eukaryotes, the enzymatic activities required for this process are supplied by a replicative DNA polymerase, Flap endonuclease 1 (Fen1) and DNA ligase 1 (Lig1). These factors interact with the sliding clamp PCNA (proliferating cell nuclear antigen) providing a potential means of co-ordinating their sequential actions within a higher order assembly. In hyperthermophilic archaea of the Sulfolobus genus, PCNA is a defined heterotrimeric assembly and each subunit interacts preferentially with specific client proteins. We have exploited this inherent asymmetry to assemble a PCNA-polymerase-Fen1-ligase complex on DNA and have visualized it by electron microscopy. Our studies reveal the structural basis of co-occupancy of a single PCNA ring by the three distinct client proteins.
- Subjects
OKAZAKI fragments; SULFOLOBUS solfataricus; DNA replication; DNA polymerases; ENZYME kinetics
- Publication
Biochemical Journal, 2015, Vol 465, Issue 2, p239
- ISSN
0264-6021
- Publication type
Article
- DOI
10.1042/BJ20141120