Descripción del proyecto
Disección del mecanismo de la reparación de lesiones del ADN
Las lesiones genómicas conocidas como entrecruzamiento cromosómico de ADN (ICL, por sus siglas en inglés) se forman por un enlace covalente entre las dos hebras opuestas del ADN. Los ICL son muy tóxicos porque pueden interferir con la replicación y transcripción del ADN y pueden dar lugar a afecciones como la anemia de Fanconi si se dejan sin reparar. El proyecto financiado con fondos europeos ICL CHROM tiene por objeto delinear el proceso de reparación del ICL e identificar las proteínas clave implicadas, incluidas las proteínas de remodelación de nucleosomas y las chaperonas de histonas. A través de técnicas novedosas, los científicos revelarán el mecanismo regulador de la reparación del ICL y la manera en que las células remodelan su cromatina para hacer frente a los ICL.
Objetivo
DNA interstrand crosslinks can arise as a by-product of cellular metabolism and, if left unrepaired, they impede DNA replication and threaten genome integrity. Faulty repair of DNA interstrand crosslinks has been linked to Fanconi anemia (FA), a disease characterized by genomic instability and cancer predisposition. The mechanisms underlying DNA interstrand crosslink repair are not fully understood, and it is likely that key regulators of this pathway have yet to be identified. Further, as the proteins involved in DNA interstrand crosslink repair have mostly been profiled using cell-free systems, the impact of chromatin states on DNA interstrand crosslink repair is poorly understood. The main aims of this project are i) to profile the entire repertoire of proteins recruited to DNA interstrand crosslinks in human cells, and ii) to uncover and characterize chromatin proteins involved in DNA interstrand crosslink repair. First, I will develop a pull-down technique to biochemically isolate chromatin surrounding DNA interstrand crosslinks and I will employ it to quantitatively characterise the full spectrum of proteins recruited to these lesions in human cells. This approach has the potential to uncover new regulators of DNA interstrand crosslink repair, including numerous proteins with a function in chromatin biology. I will, then, complement and expand this biochemistry-based strategy using a live cell imaging approach, with the aim of uncovering nucleosome remodellers and histone chaperones recruited to DNA interstrand crosslinks. Finally, the function of new regulators will be dissected using a combination of state-of-the-art microscopy and gene targeting techniques. Together, this work will provide a comprehensive picture of the proteins involved in DNA interstrand crosslinks repair and will uncover mechanisms of chromatin remodelling associated to the repair of these lesions in human cells.
Ámbito científico
Programa(s)
Régimen de financiación
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinador
DD1 4HN Dundee
Reino Unido