The proposal spans many areas, from quantum chemistry and photophysics to biological systems, and is based on the design, characterization and application of a range of nucleic acid base analog FRET-probes. The probes and generic methodology developed here will comprise a toolbox complementing X-ray crystallography and NMR, allowing researchers to investigate biomolecules at high resolution and to get information, especially regarding dynamics and at single-molecule level, currently not obtainable. The design part emphasizes on developing a) FRET-probes for high resolution distance and orientation studies, b) probes for monitoring of long distances and the single-molecule level and c) metal nanostructures to enhance FRET distances. The extensive characterization of the probes is vital for their applications: it includes their photophysics and influence on nucleic acid structure, dynamics and thermodynamic stability. Moreover, considerable efforts will be spent taking advantage of my probes applying them in investigating 1) conformational changes of DNA upon transcription factor binding, 2) details of the conformational changes in the RNA/DNA-hybrid during transcription, 3) detailed structure and dynamics in single DNA/RNA molecule optical tweezers experiments and 4) the effect that size-expanded bases have on polymerase efficiency and fidelity. A fundamental molecular understanding of these processes gives vital insight into cellular mechanisms.
Biträdande professor vid Chalmers, Chemistry and Chemical Engineering, Chemistry and Biochemistry, Physical Chemistry
Funding Chalmers participation during 2014–2017 with 3,000,000.00 SEK