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Functional genomics of transcriptional regulation in crowded media


The present proposal concerns a return and reintegration grant for Dr Silvia Zorrilla, a current Spanish EIF fellow in the Centre de Biochimie Structurale in Montpellier (France), in order to continue doing research in the Instituto de Química Física Rocasolano (CSIC, Madrid), where she carried out her doctoral thesis, with the final objective of being permanently reincorporated there. During the EIF project, biophysical studies were conducted in parallel with structural determinations and microbiological studies by our collaborators, to better understand the function of several relatively unknown regulatory proteins from Bacillus subtilis identified in a previous European grant.

In the present project we propose to go farther in the understanding of the conditional interactions of the transcriptional regulators studied during the EIF training period, by conducting biophysical experiments in macromolecular crowding conditions mimicking the crowded nature inherent to all living systems. The methodology applied during the EIF research to the study of transcriptional regulation in the model bacterium will also be extended to some PPAR nuclear receptors, a family of eukaryotic transcriptional regulators implicated in a number of major diseases including cancer. PPAR interactions with DNA and ligands will be studied in vitro, in diluted as well as in macromolecular crowding conditions, but also in vivo using state-of-the-art single molecule fluorescence spectroscopy approaches.

The proposed research will be carried out in collaboration with groups of experts from the CSIC (Spain) and the CBS (France). On a broader level, our investigation will involve the development of single-molecule fluorescence-based homogeneous assays, adapted to be performed in crowded conditions, which may be useful in the screening for bacterial cell growth inhibitors and therapeutic drugs directed against some of the major human diseases.

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