Objective
The molecular recognition of nucleic acids by proteins is central to many cellular and biochemical processes. One important type of interaction, which is poorly understood, is the contact of phosphodiester linkages in RNA and DNA to basic amino acids such as arginine, lysine and histidine. Such interactions provide both hydrogen-bonding and electrostatic components and appear to be crucial in many cases of protein-nucleic acid recognition. This research project focuses on a novel technique of chemical cross-linking (developed in Moscow) that relies on spatial proximity of a basic amino acid in a protein to an activated pyrophosphate to identify essential phosphate interactions in nucleic acids substrates. The pyrophosphate is introduced chemically into a specific location in a synthetic DNA substrate. It is planned to extend this technique to the introduction of activated pyrophosphates into synthetic RNA duplexes.
This technique will be applied to the study of RNA-protein interactions by focusing on two essential Human Immunodeficiency Virus gene regulatory proteins: tat and rev (studied in Cambridge) which both contain regions of highly basic amino acids that are crucial for recognition of their respective RNA targets (TAR and RRE). The technique will also be developed further to study the interaction of the Vsr mismatch endonuclease with its substrate DNA (studied in Göttingen) which is important in a newly discovered pathway of macromolecular DNA metabolism. Pyrophosphate-modified DNA will be useful here also to act as an enzyme inhibitor for kinetic studies of the enzyme and also for DNA-protein cross-linking studies. The two sections of the project act as general paradigms of RNA-protein and DNA-protein interactions from which fundamental principles should emerge. The Moscow laboratory has pioneered the cross-linking chemistry in DNA whilst the laboratories in Cambridge and Göttingen have specialist expertise in RNA-protein and DNA-protein interactions respectively.
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Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
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Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Call for proposal
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Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Funding Scheme
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Coordinator
CB2 2QH Cambridge
United Kingdom
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.