Binding specificity of escherichia coli single-stranded DNA binding protein for the χ subunit of DNA pol III holoenzyme and PriA helicase

Alexander G. Kozlov, Maria J. Jezewska, Wlodzimierz Bujalowski, Timothy M. Lohman

    Research output: Contribution to journalArticle

    41 Scopus citations

    Abstract

    The Escherichia coli single-stranded DNA binding protein (SSB) plays a central role in DNA metabolism through its high affinity interactions with ssDNA, as well as its interactions with numerous other proteins via its unstructured C-termini. Although SSB interacts with at least 14 other proteins, it is not understood how SSB might recruit one protein over another for a particular metabolic role. To probe the specificity of these interactions, we have used isothermal titration calorimetry to examine the thermodynamics of binding of SSB to two E. coli proteins important for DNA replication, the χ subunit of DNA polymerase III holoenzyme and the PriA helicase. We find that an SSB tetramer can bind up to four molecules of either protein primarily via interactions with the last ∼9 amino acids in the conserved SSB C-terminal tails (SSB-Ct). We observe intrinsic specificity for the binding of an isolated SSB-Ct peptide to PriA over χ due primarily to a more favorable enthalpic component. PriA and χ also bind with weaker affinity to SSB (in the absence of ssDNA) than to isolated SSB-Ct peptides, indicating an inhibitory effect of the SSB protein core. Although the binding affinity of SSB for both χ and PriA is enhanced if SSB is prebound to ssDNA, this effect is larger with PriA indicating a further enhancement of SSB specificity for PriA. These results also suggest that DNA binding proteins such as PriA, which also interact with SSB, could use this interaction to gain access to ssDNA by first interacting with the SSB C-termini.

    Original languageEnglish (US)
    Pages (from-to)3555-3566
    Number of pages12
    JournalBiochemistry
    Volume49
    Issue number17
    DOIs
    StatePublished - May 4 2010

    ASJC Scopus subject areas

    • Biochemistry

    Fingerprint Dive into the research topics of 'Binding specificity of escherichia coli single-stranded DNA binding protein for the χ subunit of DNA pol III holoenzyme and PriA helicase'. Together they form a unique fingerprint.

  • Cite this