Insights into RNA G4ome: comprehensive characterization of G-quadruplex dependent translation controlled by RNA binding proteins – G4ome

Polysome profiling, Reporter assays, RNA-protein interaction techniques , co-immunoprecipitations, bioinformatics

Our results identified specific mRNAs translationnally regulated by a G4-dependent mechanism and characterized the mechanism of action of an RBP in this translational control.

This work will offer an informative snapshot of RNA G4 biology that could support future studies on G4-RBP interactions and G4 functions in other cells and organisms, or in different biological condition. The knowledge gained during the G4ome project will shed light on how these structures impact gene expression and has implications for understanding the role of RNA G4s in disease and treatment.

Le Bras, M., Herviou, P., Hieblot, C., Gilhodes, J., Hugnot, J.P., Ameadan, A., Dassi, E., Cammas, A., Millevoi, S. (in revision) hnRNP H/F drive G-quadruplex-mediated translation linked to genomic instability and therapy resistance in glioblastoma. Nature Commun.

RNAs can form stable four-stranded G-quadruplexes (G4) structures in vitro and in vivo. Although RNA G4s have been implicated in post-transcriptional gene regulation and diseases, the G4 RNA-protein network underlying the function of these structures remains to be fully characterized. Until now, knowledge on the existence, the role and the mechanism of action of RNA G4s through RNA-binding protein (RBP) regulation has been mainly obtained by studies of specific G4-containing mRNAs. The general objective of this research proposal is to provide a comprehensive view of the regulatory function of RNA G4s in mRNA translation and the RBP associated to these structures. Through a multi-disciplinary approach including genome-wide analysis, biophysics, molecular and cellular biology, the present fundamental project aims at identifying i) functional RNA G4s regulating protein synthesis, ii) their protein partners in physiological conditions and iii) the associated regulatory molecular mechanisms in human cells. This work will offer an informative snapshot of RNA G4 biology that could support future studies on G4-RBP interactions and G4 functions in other cells and organisms, or in different biological condition. The knowledge gained during the G4ome project will shed light on how these structures impact gene expression and has implications for understanding the role of RNA G4s in disease and treatment. Besides this cognitive ambition, our high-throughput approach applied to capture RBPs assembled on defined RNA elements meet urgent need in RNA research and will provide a proof of principle model for future studies concerning other RNA elements/structures relevant for post-transcriptional regulation.