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. Programme(s) IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993- Topic(s) 36 - Biochemistry and New Pharmaceutical Compounds Call for proposal Data not available Funding Scheme Data not available Coordinator MRC Laboratory of Molecular Biology EU contribution No data Address Hills Road CB2 2QH Cambridge United Kingdom See on map Total cost No data Participants (2) Sort alphabetically Sort by EU Contribution Expand all Collapse all Georg-August Universität of Göttingen Germany EU contribution No data Address 37077 Göttingen See on map Total cost No data Moscow State University M.V. Lomonosov Russia EU contribution No data Address 119899 Moscow See on map Total cost No data
