Role of PCNA and TLS polymerases in D-loop extension during homologous recombination in humans |
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| Authors: | Marek Sebesta Peter Burkovics Szilvia Juhasz Sufang Zhang Judit E. Szabo Marietta Y.W.T. Lee Lajos Haracska Lumir Krejci |
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| Affiliation: | 1. National Centre for Biomolecular Research, Masaryk University, Brno, Czech Republic;2. Department of Biology, Masaryk University, Brno, Czech Republic;3. International Clinical Research Center, Center for Biomolecular and Cellular Engineering, St. Anne''s University Hospital Brno, Brno, Czech Republic;4. Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary;5. Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY, USA;6. Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary |
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| Abstract: | Homologous recombination (HR) is essential for maintaining genomic integrity, which is challenged by a wide variety of potentially lethal DNA lesions. Regardless of the damage type, recombination is known to proceed by RAD51-mediated D-loop formation, followed by DNA repair synthesis. Nevertheless, the participating polymerases and extension mechanism are not well characterized. Here, we present a reconstitution of this step using purified human proteins. In addition to Pol δ, TLS polymerases, including Pol η and Pol κ, also can extend D-loops. In vivo characterization reveals that Pol η and Pol κ are involved in redundant pathways for HR. In addition, the presence of PCNA on the D-loop regulates the length of the extension tracks by recruiting various polymerases and might present a regulatory point for the various recombination outcomes. |
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| Keywords: | TLS polymerases Homologous recombination DNA repair synthesis D-loop Reconstitution |
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