GLORI · Global analysis of R-loop structures (RNA-DNA hybrids) by advanced microscopic and genetic approaches
FP7 — People (Marie Curie Actions)
- Duration
- 2012-12-01 → 2016-11-30
- EU contribution
- €100,000
- Participants
- 1
- Scheme
- MC-CIG
Lines connect the coordinator with its partners.
Results in brief
Global analysis of R-loop structures (RNA-DNA hybrids) by advanced microscopic and genetic approaches
Chromosomal breaks are usually associated with pathological disorders but a growing body of evidence point towards their role in a number of physiological processes, as meiotic recombination or class switch recombination. The main goal of this project was three-fold: first, we aimed to find associations between the occurrence of certain histone modifications / histone modifying enzymes, non-B DNA structures (e.g. RNA-DNA hybrids) and DNA damage formation (as DNA double-strand breaks (DSBs) and single-strand breaks (nicks). Second, we wished to understand how the molecular components of DNA breakage - associated or no with RNA-DNA hybrid formation - is embedded with higher-order chromatin architecture. Third, we wished to characterize protein factors that are involved in sensing, forming, stabilizing, and eliminating scheduled and unscheduled R-loop structures. Our investigations were performed in three model organisms, from S. cerevisiae to human cell lines, upon various metabolic settings and in mitotic and meiotic growth conditions.
Data: CORDIS, © European Union
Project objective
Genetic alterations from point mutations to chromosome rearrangements are usually associated with pathological disorders. Unraveling the origin of such events was greatly enhanced by the discovery of cis-acting factors (e.g. chromosomal fragile sites, G quadruplexes)behaving as chromosomal hotspots for genomic instability, but a detailed mechanistic understanding of their mode of action has remained elusive. Most current models of genomic instability incorporate R-loops - nucleic acid structures in which an RNA strand displaces onestrand of a duplex DNA molecule for a limited length - as immediate cause of gene rearrangements. The present application is based on prior observations of the host laboratory on previously unrecognized ribonucleoprotein particles containing RNA-DNA hybrids, being positioned at loop-size intervals in the chromatin of lower and higher eukaryotic cells (Szekvolgyi L et al., PNAS, 2007). The molecular identity and the cell biological significance of the revealed structures are unknown. The goal of this project is to unravel the molecular structure and epigenetic landscape of the regularly spaced ribonucleoprotein complexes containing RNA-DNA hybrids previously described by our laboratory, with a special emphasis on their connection to chromatin loop domain formation and genetic recombination. We expect that our work plan - proposing the global analysis of R-loop structures using a range of cell biophysical and genetic approaches in the genetically tractable model organism S cerevisiae as well as in various human cell lines - will yield novel data relevant to the rules that govern not only normal chromatin architecture but also an RNA-DNA hybrid-mediated pathological scenario leading to chromosomalrearrangements.
Original text from CORDIS.
Participants
- DEBRECENI EGYETEM · DebrecenCoordinatorHungary
Links
Data: CORDIS, © European Union