Description du projet
Comprendre la façon dont la nature contourne les obstacles à la transcription
Les lésions de l’ADN bloquant la transcription peuvent être déclenchées par une grande variété d’agents endommageant l’ADN et constituent un obstacle pour les enzymes de transcription des gènes. La réparation couplée à la transcription (TCR) est une voie spécialisée de réparation de l’ADN qui élimine sélectivement les lésions de l’ADN des gènes activement transcrits, rétablissant ainsi la transcription. Les mécanismes qui sous‑tendent ces processus sont mal compris. Le projet STOP‑FIX‑GO, financé par l’UE, permettra d’élucider les mécanismes qui sous‑tendent les réponses cellulaires aux dommages qui bloquent la transcription, grâce à une combinaison de techniques de génomique, de protéomique et de cartographie de haute technologie, en se concentrant sur les prometteuses protéines TCR. Comprendre comment les cellules surmontent naturellement les obstacles à la transcription pourrait déboucher sur de nouveaux traitements.
Objectif
Bulky DNA lesions are a major obstacle during gene transcription by RNA polymerase II enzymes (RNAPII). The stalling of RNAPII at DNA lesions triggers a genome-wide transcriptional arrest. Transcription-coupled repair (TCR) is a specialized DNA repair pathway that selectively removes DNA lesions from actively transcribed genes to restore transcription. Stalled RNAPII at DNA lesions forms a roadblock for advancing DNA replication forks resulting in toxic collisions. The mechanisms that enable the repair of transcription-blocking DNA lesions, the restoration of transcription after repair and the resolution of transcription-replication conflicts are poorly understood. To address these knowledge gaps, I propose to establish a series of innovative approaches aimed at identifying the mechanisms involved in the cellular responses to transcription-blocking DNA damage. We will focus on the functional characterization of known and several promising new TCR factors that we recently identified in combined genome-wide CRISPR and targeted proteomics screens. I propose to dissect the role of known and new TCR proteins by (1) applying a genome-wide approach for directly measuring TCR activity in combination with proximity-labelling proteomics and genetic-interaction mapping to define how TCR complexes assemble and operate, (2) identifying the mechanisms in transcription restoration by combining advanced genomics methods to map nascent transcripts, monitor RNAPII occupancy, and correlate these with specific chromatin modifications in a genome-wide manner, and (3) dissecting the mechanisms involved in resolving transcription-replication conflicts by combining functional DNA replication assays with genome-wide approaches to map transcription, R-loops and DNA replication directionality. This ERC project will break new grounds by offering a detailed understanding of the mechanisms that enable cells to overcome transcriptional roadblocks.
Champ scientifique
Programme(s)
- HORIZON.1.1 - European Research Council (ERC) Main Programme
Régime de financement
ERC - Support for frontier research (ERC)Institution d’accueil
2333 ZA Leiden
Pays-Bas