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Electron microscopic analysis reveals that replication factor C is sequestered by single-stranded DNA.

R C Keller·R Mossi·G Maga·R E Wellinger·U Hübscher·J M Sogo
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Abstract

Replication factor C (RF-C) is a eukaryotic heteropentameric protein required for DNA replication and repair processes by loading proliferating cell nuclear antigen (PCNA) onto DNA in an ATP-dependent manner. Prior to loading PCNA, RF-C binds to DNA. This binding is thought to be restricted to a specific DNA structure, namely to a primer/template junction. Using the electron microscope we have examined the affinity of human heteropentameric RF-C and the DNA-binding region within the large subunit of RF-C from Drosophila melanogaster (dRF-Cp140) to heteroduplex DNA. The electron microscopic data indicate that both human heteropentameric RF-C and the DNA-binding region within dRF-Cp140 are sequestered by single-stranded DNA. No preferential affinity for the 3' or 5' transition points from single- to double-stranded DNA was evident.

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AnimalsDNA, Single-StrandedDNA-Binding ProteinsDrosophilaHeLa CellsHomeodomain ProteinsHumansMicroscopy, ElectronMinor Histocompatibility AntigensNucleic Acid HeteroduplexesPlasmidsProtein BindingProto-Oncogene Proteins c-bcl-2Replication Protein CRepressor ProteinsSaccharomyces cerevisiae ProteinsJournal ArticleResearch Support, Non-U.S. Gov't

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R C Keller, R Mossi, G Maga, R E Wellinger, U Hübscher, J M Sogo. 1999-09-01. Electron microscopic analysis reveals that replication factor C is sequestered by single-stranded DNA.. https://doi.org/10.1093/nar%2F27.17.3433

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