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Understanding the functions of the BRCA protein complexes in DNA break repair and in replication stress

Project description

Role of BRCA protein complexes in DNA double-strand break repair and in replication fork remodelling

BRCA1 and BRCA2 genes and associated factors are often mutated in breast and ovarian cancers. BRCA proteins function together with the RAD51 recombinase in DNA double-strand break repair but have separate roles in the metabolism of challenged replication forks. The EU-funded BRCA INSIGHTS project will use a biochemical approach to study the precise functions of these proteins. The objective is to define how BRCA proteins are involved in the initial steps of DNA double-strand break repair, including nucleolytic processing of DNA breaks, RAD51 loading on ssDNA and invasion of homologous DNA template. The study aims to determine the specific separate functions of RAD51, RAD52 and BRCA1/2 complexes in fork remodelling, protection and restart, which prevent pathological degradation of nascent DNA under replication stress.

Objective

BRCA1, BRCA2 and associated factors are frequently mutated in familial and sporadic cancers of the breast and ovary. The BRCA proteins have canonical functions together with the RAD51 recombinase in DNA double-strand break repair by homologous recombination. More recently, it was discovered that the same proteins have separate, recombination-independent roles in the metabolism of challenged replication forks. It is currently debated whether defects in the canonical and/or non-canonical functions of BRCA contribute to tumorigenesis.

As BRCA1 and BRCA2 function in various processes, interact with numbers of co-factors, and can be extensively post-translationally modified, phenotypes associated with their defects are often very pleiotropic. Here I propose to use a biochemical approach, which will allow us to study the precise functions of these proteins with defined substrates and co-factors in controlled systems. BRCA1 and BRCA2 represent some of the largest factors in human proteome. Using state-of-the-art protein expression strategies, we are now able to produce sufficient quantities of the respective complexes in their native or post-translationally modified states to make biochemistry feasible.

We aim to define how the BRCA proteins help couple the initial steps in DNA break repair including nucleolytic processing of DNA breaks, RAD51 loading on ssDNA and subsequent invasion of homologous DNA template. We will also determine the precise non-canonical functions of RAD51, RAD52 and BRCA1/2 complexes in fork remodeling, fork protection and fork restart, which prevents pathological degradation of nascent DNA under replication stress. We anticipate that these diverse functions of BRCA proteins are governed by their co-factors, post-translational modifications and specific substrates. A biochemical approach is uniquely suited to understand the mechanisms how BRCA proteins regulate DNA metabolic pathways to promote genome stability and prevent tumorigenesis.

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Host institution

FONDAZIONE PER L'ISTITUTO DI RICERCA IN BIOMEDICINA
Net EU contribution
€ 1 868 152,50
Address
VIA FRANCESCO CHIESA 5
6500 Bellinzona
Switzerland

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Region
Schweiz/Suisse/Svizzera Ticino Ticino
Activity type
Research Organisations
Links
Total cost
€ 1 868 152,50

Beneficiaries (1)