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A Postincision-Deficient TFIIH Causes Replication Fork Breakage and Uncovers Alternative Rad51- or Pol32-Mediated Restart Mechanisms

Reference: Moriel-Carretero & Aguilera; Molecular Cell 37, 690-701, March 12, 2010 A Postincision-Deficient TFIIH Causes Replication Fork Breakage and Uncovers Alternative Rad51- or Pol32-Mediated Restart Mechanisms
Authors: María Moriel-Carretero* and Andrés Aguilera*

*Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla-CSIC, Sevilla, Spain
Summary: Homologous recombination is a major double-strand break (DSB) repair mechanism that acts during the S and G2 phases. In contrast, nucleotide excision repair (NER) is a major pathway for the repair of DNA bulky adducts that is unrelated to replication. We show that replication can be strongly disturbed in a specific type of rad3/XPD NER mutants of TFIIH, causing replication fork breakage. In contrast to classical NER-deficient mutations, the S. cerevisiae rad3-102 allele, which has a minimal impact on UV resistance, channels bulky adducts into DSBs. rad3-102 allow Rad1/XPF and Rad2/XPG-catalyzed DNA incisions but fail to perform post-incision steps retaining TFIIH at the damaged site. Broken forks are rescued by MRX-Rad52-Rfc1-dependent recombination via two types of replication re-start mechanisms, one being Rad51-dependent and the other Pol32-dependent. Our results define the genetic and molecular hallmarks of replication-fork breakage and re-start and bring new insights to understand specific NER-related human syndromes.
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REFERENCIA DEL GRUPO Y/O INVESTIGADOR
El trabajo del grupo dirigido por Andrés Aguilera en CABIMER se centra en entender las causas y consecuencias de la inestabilidad genómica y su relación con otros procesos biológicos tanto en los modelos S. cerevisiae y C. elegans como en cultivos celulares humanos. Trata de entender los fenómenos de inestabilidad asociados a transcripción y a replicación, el acoplamiento entre la transcripción con el transporte del ARN, y la importancia del ARN en la dinámica de los genomas. El estudio de la base genética y molecular de la enfermedad genética Xeroderma pigmentosum, un paradigma de la relación causa-efecto entre inestabilidad genética y cáncer, constituye una de las líneas específicas de investigación, dentro de la cual se ha desarrollado el trabajo publicado en Molecular Cell.


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