Project acronym BrokenGenome
Project Breaking and rebuilding the genome: mechanistic rules for the dangerous game of sex.
Researcher (PI) Corentin CLAEYS BOUUAERT
Host Institution (HI) UNIVERSITE CATHOLIQUE DE LOUVAIN
Call Details Starting Grant (StG), LS1, ERC-2018-STG
Summary Sexual reproduction depends on the programmed induction of DNA double-strand breaks (DSBs) and their ensuing repair by homologous recombination. This complex process is essential for sexual reproduction because it ultimately allows the pairing and separation of homologous chromosomes during formation of haploid gametes. Although meiotic recombination has been investigated for decades, many of the underlying molecular processes remain unclear, largely due to the lack of biochemical studies. I have recently made important progress by, for the first time, successfully purifying proteins involved in two aspects of meiotic recombination: DSB formation and the final stage of formation of the crossovers that are a central raison-d’être of meiotic recombination. This has opened new avenues for future research that I intend to pursue in my own laboratory. Here, I propose a set of biochemical approaches, complemented by molecular genetics methods, to gain insights into four central problems: (i) How meiotic proteins collaborate to induce DSBs; (ii) How DSB proteins interact with components that form the axes of meiotic chromosomes; (iii) How proteins involved at later stages of recombination form crossovers; and (iv) How crossover proteins interact with components of synapsed chromosomes. For each problem, I will set up in vitro systems to probe the activities of the players involved, their interactions with DNA, and their assembly into macromolecular complexes. In addition, I propose to develop new methodology for identifying proteins that are associated with DNA that has undergone recombination-related DNA synthesis. My goal is to gain insights into the mechanisms that govern meiotic recombination. Importantly, these mechanisms are intimately linked not only to gamete formation, but also to the general recombination pathways that all cells use to maintain genome stability. In both contexts, our findings will be relevant to the development and avoidance of disease states.
Summary
Sexual reproduction depends on the programmed induction of DNA double-strand breaks (DSBs) and their ensuing repair by homologous recombination. This complex process is essential for sexual reproduction because it ultimately allows the pairing and separation of homologous chromosomes during formation of haploid gametes. Although meiotic recombination has been investigated for decades, many of the underlying molecular processes remain unclear, largely due to the lack of biochemical studies. I have recently made important progress by, for the first time, successfully purifying proteins involved in two aspects of meiotic recombination: DSB formation and the final stage of formation of the crossovers that are a central raison-d’être of meiotic recombination. This has opened new avenues for future research that I intend to pursue in my own laboratory. Here, I propose a set of biochemical approaches, complemented by molecular genetics methods, to gain insights into four central problems: (i) How meiotic proteins collaborate to induce DSBs; (ii) How DSB proteins interact with components that form the axes of meiotic chromosomes; (iii) How proteins involved at later stages of recombination form crossovers; and (iv) How crossover proteins interact with components of synapsed chromosomes. For each problem, I will set up in vitro systems to probe the activities of the players involved, their interactions with DNA, and their assembly into macromolecular complexes. In addition, I propose to develop new methodology for identifying proteins that are associated with DNA that has undergone recombination-related DNA synthesis. My goal is to gain insights into the mechanisms that govern meiotic recombination. Importantly, these mechanisms are intimately linked not only to gamete formation, but also to the general recombination pathways that all cells use to maintain genome stability. In both contexts, our findings will be relevant to the development and avoidance of disease states.
Max ERC Funding
1 499 075 €
Duration
Start date: 2019-07-01, End date: 2024-06-30
Project acronym RESEAL
Project Epithelial Resealing
Researcher (PI) Antonio Alfredo Coelho Jacinto
Host Institution (HI) FUNDACAO CALOUSTE GULBENKIAN
Call Details Starting Grant (StG), LS1, ERC-2007-StG
Summary Epithelia have the essential role of acting as a barrier that protects living organisms and its organs from the surrounding milieu. Therefore, it is crucial for epithelial tissues to have robust ways of maintaining its integrity despite the frequent damage caused by normal cell turnover, inflammation and injury. All epithelia have some capacity to repair themselves, however, the wound-healing process differs dramatically between the developmental stage and type of tissue involved. In this project we will focus on investigating the capacity that several simple epithelial tissues have to reseal small discontinuities very rapidly and efficiently. This repair mechanism that we call epithelial resealing is based on the contraction of an actomyosin purse string in the leading edge cells around the wound margin. Epithelial resealing seems to be a fundamental repair mechanism, acting in several types of simple embryonic and adult epithelia, in both vertebrates and invertebrates. The cell biology of epithelial resealing has started to be understood but there are still many open questions and the signalling cascades that regulate this process are largely unknown. We propose to investigate epithelial resealing using a combination of genetics and high resolution live imaging. The Drosophila embryonic epithelium will be our primary model system and we will start by characterizing in detail novel genes involved in resealing that have been identified in a pilot screen previously performed in the laboratory. We will also perform a new RNAi genetic screen based on a very large collections of transgenic lines to completely unravel the signalling network that controls epithelial resealing. In order to investigate how conserved in vertebrates are the epithelial resealing mechanisms, we will establish epithelial wounding assays in zebrafish simple epithelial tissues and we will study, in this vertebrate model system, the molecular mechanisms that we will uncover using Drosophila.
Summary
Epithelia have the essential role of acting as a barrier that protects living organisms and its organs from the surrounding milieu. Therefore, it is crucial for epithelial tissues to have robust ways of maintaining its integrity despite the frequent damage caused by normal cell turnover, inflammation and injury. All epithelia have some capacity to repair themselves, however, the wound-healing process differs dramatically between the developmental stage and type of tissue involved. In this project we will focus on investigating the capacity that several simple epithelial tissues have to reseal small discontinuities very rapidly and efficiently. This repair mechanism that we call epithelial resealing is based on the contraction of an actomyosin purse string in the leading edge cells around the wound margin. Epithelial resealing seems to be a fundamental repair mechanism, acting in several types of simple embryonic and adult epithelia, in both vertebrates and invertebrates. The cell biology of epithelial resealing has started to be understood but there are still many open questions and the signalling cascades that regulate this process are largely unknown. We propose to investigate epithelial resealing using a combination of genetics and high resolution live imaging. The Drosophila embryonic epithelium will be our primary model system and we will start by characterizing in detail novel genes involved in resealing that have been identified in a pilot screen previously performed in the laboratory. We will also perform a new RNAi genetic screen based on a very large collections of transgenic lines to completely unravel the signalling network that controls epithelial resealing. In order to investigate how conserved in vertebrates are the epithelial resealing mechanisms, we will establish epithelial wounding assays in zebrafish simple epithelial tissues and we will study, in this vertebrate model system, the molecular mechanisms that we will uncover using Drosophila.
Max ERC Funding
1 150 000 €
Duration
Start date: 2008-11-01, End date: 2014-10-31
