Objective
The discovery of new anti-microbial agents has become a problem of crucial importance in light of the acquired resistance of many pathogens to most of the available antibiotics. The new industrial trend in antibacterial R&D is focused on the high through-put screening of select, appropriately chosen, broad spectrum macromolecular targets. In the present programme, we plan to develop a new class of antibiotics directed against bacterial translation initiation factor IF2 or interfering with the interaction of this protein with its main ligand initiator fMet-tRNA, which, unlike elongator tRNAs, is not present in eukaryotes. In fact, IF2 and the IF2-fMet-tRNA complex are endowed with all the properties required by an ideal target for rational drug design. High through-put screening tests will be applied in the search for both natural and synthetic compounds interfering with the various functions of IF2. The positive hits will be subjected to secondary screens and the most promising compounds will be improved by combinatorial chemistry and rational drug design based on the target's 3D structure which, in the meantime, will be elucidated by NMR spectroscopy and X-ray crystallography. Indeed, one of the main aims of this proposal is to characterize the 3D structure of initiation factor IF2, both free and in complex with its ligands (fMet-tRNA and guanosine nucleotides); more precisely, we envisage the elucidation of both crystal and solution structure of the two domains which contain all the IF2 activities required for translation (i.e. the central G-domain containing GTP binding site, GTPase catalytic center, 30S and 50S binding sites and the C-terminal domain containing the fMet-tRNA binding site). The perspectives for elucidation of the 3D structure of IF2, both alone and complexed with some of its ligands, are good. In fact, important progress in this direction has been made recently with the production of co-crystals of Bacillus stearothermophilus IF2 C-domain-fMet-tRNA complex and of good quality crystals of the G-C-domains. The former crystals diffract with close to 3.0 A resolution and native diffraction data sets as well as different heavy atom derivatives enabling phase determination by different techniques are already available. Furthermore, good quality 2D NMR spectra of 15N-labeled IF2 C-domain have been obtained and a promising screening test to select for IF2 mutants defective in fMet-tRNA binding has been worked out. An important aspect of our proposal is that it places a great emphasis on the study of RNA-protein interaction, a topic about which relatively little is known compared to protein-protein and protein-DNA interactions. The results obtained will not only enable us to clarify the molecular nature and the spatial organization of the active sites of IF2, thus enabling us to shed more light on the mechanism and regulation of translation initiation, but will also contribute a better understanding of the architecture and mode of action of the important superfamily of G-proteins of which IF2 is a member.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- medical and health sciencesbasic medicinemedicinal chemistry
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesphysical sciencesopticsspectroscopyabsorption spectroscopy
- natural sciencesbiological sciencesgeneticsnucleotides
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Call for proposal
Data not availableFunding Scheme
CSC - Cost-sharing contractsCoordinator
62032 Camerino
Italy