TAILIEUCHUNG - Báo cáo khoa học: Effect of monovalent cations and G-quadruplex structures on the outcome of intramolecular homologous recombination

Homologous recombination is a very important cellular process, as it pro-vides a major pathway for the repair of DNA double-strand breaks. This complex process is affected by many factors within cells. Here, we have studied the effect of monovalent cations (K + ,Na + , and NH4 + ) on the outcome of recombination events, as their presence affects the biochemical activities of the proteins involved in recombination as well as the structure of DNA. | ỊFEBS Journal Effect of monovalent cations and G-quadruplex structures on the outcome of intramolecular homologous recombination Paula Barros Francisco Boan Miguel G. Blanco and Jaime Gomez-Marquez Departamento de Bioquimica e Bioloxia Molecular Facultade de Bioloxia-CIBUS Universidade de Santiago de Compostela Spain Keywords G-quadruplex minisatellite MsH43 monovalent cations recombination fidelity repetitive sequences Correspondence J. Gomez-Marquez Departamento de Bioquimica e Bioloxia Molecular Facultade de Bioloxia-CIBUS Universidade de Santiago de Compostela 15782 Santiago de Compostela Spain Fax 34 9815969054 Tel 34 981563100 ext. 16937 E-mail These authors contributed equally to this work Received 17 February 2009 revised 18 March 2009 accepted 20 March 2009 doi Homologous recombination is a very important cellular process as it provides a major pathway for the repair of DNA double-strand breaks. This complex process is affected by many factors within cells. Here we have studied the effect of monovalent cations K Na and NH4 on the outcome of recombination events as their presence affects the biochemical activities of the proteins involved in recombination as well as the structure of DNA. For this purpose we used an in vitro recombination system that includes a protein nuclear extract as a source of recombination machinery and two plasmids as substrates for intramolecular homologous recombination each with two copies of different alleles of the human minisatellite MsH43. We found that the presence of monovalent cations induced a decrease in the recombination frequency accompanied by an increase in the fidelity of the recombination. Moreover there is an emerging consensus that secondary structures of DNA have the potential to induce genomic instability. Therefore we analyzed the effect of the sequences capable of forming G-quadruplex on the production of recombinant molecules taking advantage of the

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