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A Single‐Molecule Reconstitution of Translesion Synthesis and Competition Between DNA Polymerases
Author(s) -
Kath James,
Jergic Slobodan,
Heltzel Justin,
Jacob Deena,
Dixon Nicholas,
Sutton Mark,
Walker Graham,
Loparo Joseph
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.561.6
Subject(s) - processivity , dna polymerase , polymerase , dna polymerase ii , dna clamp , dna polymerase delta , dna replication , dna polymerase mu , biology , microbiology and biotechnology , chemistry , dna , genetics , circular bacterial chromosome , reverse transcriptase , polymerase chain reaction , gene
Translesion synthesis (TLS) alleviates replication stalling at DNA lesions. Ring‐shaped processivity clamps play a critical but ill‐defined role in mediating exchange between replicative and TLS DNA polymerases at sites of DNA damage. We have developed a single‐molecule approach to reconstitute TLS and exchange of the Escherichia coli replicative polymerase, Pol III, with the TLS polymerases Pol II and Pol IV. We observe clamp‐mediated exchange of Pol III with both TLS polymerases, and that distinct sets of interactions of each polymerase with the clamp govern the mode of exchange. These results suggest polymerase selection is not simply governed by a mass action competition, but also by positioning of polymerases on clamps at replication intermediates, suggesting a hierarchy for access to DNA lesions.