Objective Thiamine pyrophosphate (TPP) is the active form of vitamin B1. As well as being an important cofactor for primary metabolism, TPP also controls gene regulation through riboswitches in bacteria, fungi, plants and algae. Riboswitches are evolutionarily conserved sequences in the mRNA to which a metabolite binds directly, thereby affecting expression of the protein. In the green alga Chlamydomonas reinhardtii, riboswitches have been found to control gene expression of two thiamine genes, THIC and THI4. To date, studies on riboswitches remain descriptive. Here we propose to investigate, for the first time, in vivo, the role of riboswitches in regulating biosynthetic pathways, using the thiamine pathway in C. reinhardtii. This will be achieved by analyzing the vitamin B1 metabolism of THIC mutants transformed with a functional THIC gene with or without its riboswitch. Additionally, TPP riboswitches will be introduced into yeast, which lacks riboswitches. Data will be used to build a mathematical model of thiamine metabolism with or without riboswitches. We also plan to look for new ‘TPP riboswitch-controlled genes’ by differential display proteomics. This will also give us the opportunity to create a promising molecular tool for the fine-tuning of gene expression. As a proof of concept, a TPP riboswitch will be fused to a reporter gene and transformed either into the nucleus or into the chloroplast of C. reinhardtii. This will allow us to manipulate gene expression and investigate gene function by adding vitamin B1 to the medium. The research project will bring new insight to our understanding of regulation of vitamin metabolism. This will facilitate our ability to manipulate vitamin levels in plants thereby improving their nutritional quality. Understanding vitamin and cofactor metabolism in algae is also essential for the production of third generation biofuels, which are highly dependent on vitamin metabolism. Fields of science natural sciencesbiological sciencesmolecular biologymolecular geneticsnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics Programme(s) FP7-PEOPLE - Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Topic(s) FP7-PEOPLE-2009-IEF - Marie Curie Action: "Intra-European Fellowships for Career Development" Call for proposal FP7-PEOPLE-2009-IEF See other projects for this call Funding Scheme MC-IEF - Intra-European Fellowships (IEF) Coordinator UNIVERSITE DE GENEVE EU contribution € 173 065,20 Address RUE DU GENERAL DUFOUR 24 1211 Geneve Switzerland See on map Region Schweiz/Suisse/Svizzera Région lémanique Genève Activity type Higher or Secondary Education Establishments Administrative Contact Teresa Fitzpatrick (Prof.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data
