Project description DEENESFRITPL Mechanistic insight into post-translational modifications of proteins Protein function, stability and interaction with other proteins is often regulated by post-translational modifications. Ubiquitination is a post-translational modification process that involves the covalent attachment of ubiquitin molecules to target proteins by enzymes called E3 ubiquitin ligases. Ubiquitination plays a crucial role in maintaining protein homeostasis by facilitating the efficient elimination of damaged or unnecessary proteins. Funded by the European Research Council, the RINGE3 project aims to investigate how RING E3 enzymes interact with other ubiquitin-conjugating enzymes (E2) and substrates, and how they are regulated. The results will help us better understand the mechanisms of RING E3-mediated ubiquitination and may lead to new therapies for diseases such as cancer. Show the project objective Hide the project objective Objective Ubiquitin (Ub) conjugation regulates a myriad of cellular processes in the eukaryotic cell. Ub-ligases (E3) play a pivotal role in deciding the substrate’s fate and function by catalyzing the transfer of Ub from Ub-conjugating enzyme (E2) to a substrate protein lysine sidechain. Successive rounds of E3-catalyzed substrate ubiquitination lead to the formation of poly-Ub chains or multi-monoubiquitination, which direct the substrate to different biological fates such as degradation by the 26S proteasome. RING E3s comprise the largest family of E3s with approximately 600 members in humans. Over the last fifteen years, structural biology and biochemical studies have paved the way for understanding how RING E3s interact with E2s and substrates, and how they are regulated. Recently my group has trapped the crystal structure of a RING E3 bound to an E2 covalently-linked to Ub (E2~Ub), thus providing a molecular snapshot of how RING E3 optimizes E2~Ub for catalysis. Despite these advances, the mechanisms of RING E3-catalyzed ubiquitination are not completely understood. Here, we propose to investigate three key aspects of RING E3 functions. First, we will determine structures of several RING E3s bound to E2~Ub to dissect the molecular basis for RING E3-E2~Ub selectivity. Second, our recent structure of a RING E3, Cbl-b, bound to E2~Ub and a substrate peptide provides a starting point for structural determination of a more challenging RING E3-E2~Ub-intact substrate complex to elucidate the mechanisms of substrate ubiquitination. Third, we have developed an ubiquitinated Cbl-substrate mimetic to study the mechanisms of RING E3-catalyzed poly-ubiquitination using structural and biochemical approaches. Expected results will greatly expand our knowledge of RING E3-mediated ubiquitination and will foster strategies in exploiting E3s for therapeutic development, since deregulation of E3s underlies many diseases including cancers. Fields of science natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomicsnatural scienceschemical sciencescatalysisnatural scienceschemical sciencesorganic chemistryaminesnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymesnatural sciencesbiological sciencesmolecular biologystructural biology Programme(s) H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme Topic(s) ERC-CoG-2014 - ERC Consolidator Grant Call for proposal ERC-2014-CoG See other projects for this call Funding Scheme ERC-COG - Consolidator Grant Host institution BEATSON INSTITUTE FOR CANCER RESEARCH LBG Net EU contribution € 2 000 000,00 Address SWITCHBACK ROAD GARSCUBE ESTATE G61 1BD Bearsden United Kingdom See on map Region Scotland West Central Scotland East Dunbartonshire Activity type Research Organisations Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 2 000 000,00 Beneficiaries (1) Sort alphabetically Sort by Net EU contribution Expand all Collapse all BEATSON INSTITUTE FOR CANCER RESEARCH LBG United Kingdom Net EU contribution € 2 000 000,00 Address SWITCHBACK ROAD GARSCUBE ESTATE G61 1BD Bearsden See on map Region Scotland West Central Scotland East Dunbartonshire Activity type Research Organisations Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 2 000 000,00
