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

Keller, Robert C. ; Mossi, Romina ; Maga, Giovanni ; Wellinger, Ralf E. ; Hubscher, Ulrich ; Sogo, José M.

In: Nucleic Acids Research, 1999, vol. 27, no. 17, p. 3433-3437

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    Title
    • Electron microscopic analysis reveals that replication factor C is sequestered by single-stranded DNA
    Author
    • Keller, Robert C.. Institute for Cell Biology, ETH-Hönggerberg, CH-8049 Zürich, Switzerland
    • Mossi, Romina. Department of Veterinary Biochemistry, University Zürich-lrchel, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland
    • Maga, Giovanni. Institute of Biochemical and Evolutionary Genetics IGBE-CNR, National Research Council, via Abbiategrasso 207, Pavia 1-27100, Italy
    • Wellinger, Ralf E.. Institute for Cell Biology, ETH-Hönggerberg, CH-8049 Zürich, Switzerland
    • Hubscher, Ulrich. Department of Veterinary Biochemistry, University Zürich-lrchel, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland
    • Sogo, José M.. Institute for Cell Biology, ETH-Hönggerberg, CH-8049 Zürich, Switzerland, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland, via Abbiategrasso 207, Pavia 1-27100, Italy
    Document Type
    • Postprint
    Language
    • English
    Published in
    • Nucleic Acids Research, 1999, vol. 27, no. 17, p. 3433-3437. Oxford University Press
    Other Version
    Classification
    • Health
    OAI-PMH Identifier
    • oai:doc.rero.ch:301429
    Summary
    Replication factor C (RF-C) is a eukaryotic hetero-pentameric 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