Analysis of the processing of complex recombination intermediates by structure-selective endonucleases

  1. Raquel Carreira Rodríguez
Dirixida por:
  1. Miguel González Blanco Director

Universidade de defensa: Universidade de Santiago de Compostela

Fecha de defensa: 10 de marzo de 2023

  1. Dana Branzei Presidente/a
  2. Anxo Vidal Figueroa Secretario
  3. Aura Carreira Moreno Vogal
  1. Departamento de Bioquímica e Bioloxía Molecular

Tipo: Tese


DNA repair by homologous recombination involves the formation of branched intermediates that can constitute a source of genome instability when untimely processed. To prevent these structures from interfering with chromosome segregation, cells rely on the action of the structure-selective endonucleases Mus81-Mms4 and Yen1. Here, we employed a biochemical approach to show that both endonucleases can process the central intermediate of this pathway, the displacement loop (D-loop). Using synthetic and enzymatically reconstituted D-loops, we mapped their incision sites and recapitulated for the first time the formation of a half-crossover precursor in vitro. This implies that the concurrent action of Mus81-Mms4 and Yen1 on a D-loop may lead to the generation of complex chromosomal rearrangements.