Working Group 2 – Mineralocorticoid Receptor Structure and Function
Working Group 2 Members
Associate Professor, Department of Physiology and Institute of Biomedical Technology.
University of Laguna, Spain
Keywords: Post-translational modifications, ion channels, electrophysiology, whole animal physiology
Head: Professor Brian Harvey
Director, Molecular Medicine Research Laboratories
Royal College of Surgeons in Ireland (RCSI), Beaumont Hospital, Dublin, Ireland
Keywords: Epithelial Na+ channel (ENaC), estrogen, ion channels, kidney, intestine
Research Group 5
Head: Professor Rosario González Muñiz
Research Scientist at the Medicinal Chemistry Institute (IQM-CSIC), Madrid, Spain
Keywords: Medicinal chemistry, antagonists for bioactive peptides, modulators of ionchannels, non-steroid MR ligands
A key outcome will be a deeper knowledge of the molecular MR-structure-function-relationship from exploring the influence of MR subdomains and modifications for mediating or modulating (i) classical genomic MR functions, (ii) novel genomic MR effects and (iii) non-genomic MR-protein interactions.
The mineralocorticoid receptor MR is a ligand-dependent transcription factor without a strictly MR-specific hormone-response-element and with partially promiscuous ligands. Nevertheless, the MR elicits specific effects of physiological and pathological relevance in different tissues which will be characterized in detail in WGs 3-6. This WG aims to characterize the importance and consequences of different receptor domains and post-transcriptional modifications for MR function in detail. Using innovative screening approaches including ChIP-Seq experiments for identifying new MR target genes, advanced fluorescent imaging techniques for investigating protein-protein interactions and to determine subcellular spatiotemporal trafficking patterns as well as peptide arrays for investigating post-translational modifications we plan to
- Untangling the mechanism and impact of compartmentalized MR signaling at the plasma membrane and in the cytosol, ER and nucleus.
- Characterizing the influence of post-translational modifications on MR function and stability
- Assessing the effect of interaction with tissue specific co-regulators.
- Exploration of MR domains responsible for protein-protein interactions and the corresponding signaling partners as well as the spatiotemporal organization and functional consequences of these interactions.
A set of fully functional fluorescent MR mutants is available to manifest the interaction between MR structure and function. Individual interactions will be characterized by a wide range of state-of-the-art molecular biology, protein biochemistry and cell biology methods including sophisticated approaches such as FRET and BRET. Read-out parameters for MR function include DNA-binding tests (ChIP), MR transactivation hormone-response-element-reporter-gene assays, non-genomic mitogen activated kinase phosphorylation assays and co-immunoprecipitation/co-localization experiments. A variety of tagged MR-plasmids and also MR-specific antibodies for the different applications already exist.