Research objectives and content
Antibiotic producing bacteria need an effective self-defense mechanism. In the case of antibiotics acting on elongation factor Tu, the expression of specialized resistant EF-Tus may enable the producers to survive. Firstly, the mechanism of inhibition of the antibiotic GE2270A on the peptide-chain elongation cycle and its binding to the target protein EF-Tu will be examined by analyzing the resistant EF-Tu from the GE2270A-producing Planobispora rosea and mutated EF-Tus from E. coli. The results will be compared to those described for kirromycin and pulvomycin. Secondly, OtrA, the EF-G-like oxytetracyclineresistance protein of Streptomyces rimosus, will be investigated by in vitro studies in order to elucidate its functioning in ribosome reactivation. Mutant OtrA that binds XTP instead of GTP will give a clue about the individual contributions of OtrA and EF-G/EF-Tu. Altogether, the results will not only contribute to the understanding of self-defense mechanisms in antibiotic producing organisms, but also to an expansion of current views on the fundamental process of protein biosynthesis.
Training content (objective, benefit and expected impact)
The training objective of the project is to gain experiences in in vitro and in vivo translation systems, in the isolation and functional characterization of proteins and in the analysis of antibiotic inhibition mechanisms. Besides learning technical skills, I also expect to benefit for my scientific experience from the excellent research climate at the Leiden University and the group of Dr. Kraal, in particular. Furthermore, via the active participation of the group in the EU Network ERBCHRXCT 940510 I will have favourable contacts with other member groups working on translation factors with complementary approaches such as X-ray diffraction. Links with industry / industrial relevance (22)