Protein-engineering based approach for super-resolution imaging of nucleoporin-chromatin interactions

Project Information

NUPTAG

Grant agreement ID: 331373

Project closed

Start date 1 January 2014

End date 31 December 2015

Funded under

• Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)

Total cost € 161 968,80

EU contribution € 161 968,80

Coordinated by

EUROPEAN MOLECULAR BIOLOGY LABORATORY
Germany

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 (EuroSciVoc)

CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.

• natural sciences › physical sciences › optics › microscopy › super resolution microscopy
• natural sciences › biological sciences › biochemistry › biomolecules › proteins
• natural sciences › chemical sciences › organic chemistry › amines

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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
• Region: Baden-Württemberg › Karlsruhe › Heidelberg, Stadtkreis
• Activity type: Research Organisations
• Total cost: No data