Objective Nucleoporins are major building blocks of the nuclear pore complex (NPC). In addition to their role in the nucleocytoplasmic transport, a subset of nucleoporins have additional gene regulatory roles. They can bind chromatin and chromatin modifiers, either directly at the NPC or in the nucleoplasm, leading to gene repression or activation. Due to a lack of techniques that allow mapping spatial and temporal heterogeneities of individual proteins with high precision, the molecular mechanisms underlying these phenomena are far from being understood. To overcome this, I will develop a combined protein-engineering/super-resolution approach for single amino acid targeted labelling and tracking of proteins inside the cells. This strategy is based on the translational incorporation of newly developed non-canonical amino acids (ncAAs). These ncAAs are suitable for inverse-electron-demand Diels-Alder reaction, a special type of highly selective and rapid “click-chemistry” reactions proceeding in a non-toxic and fully biocompatible way inside living cells. I will employ this technology to systematically label a set of nucleoporins and chromatin modifiers with small dyes. The high photostability of these fluorophores permits long-term imaging and will allow me to identify nucleoporin binding partners in cultured mammalian cells with nanometer resolution. This will set the stage for a technology that enables deciphering protein plasticity in vivo at the molecular level and provide novel insights into multifunctionality of nucleoporins. Fields of science natural sciencesphysical sciencesopticsmicroscopysuper resolution microscopynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural scienceschemical sciencesorganic chemistryamines 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-2012-IEF - Marie-Curie Action: "Intra-European fellowships for career development" Call for proposal FP7-PEOPLE-2012-IEF See other projects for this call Funding Scheme MC-IEF - Intra-European Fellowships (IEF) Coordinator EUROPEAN MOLECULAR BIOLOGY LABORATORY EU contribution € 161 968,80 Address Meyerhofstrasse 1 69117 Heidelberg Germany See on map Region Baden-Württemberg Karlsruhe Heidelberg, Stadtkreis Activity type Research Organisations Administrative Contact Jillian Rowe (Ms.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data