Odborný seminář: Single Molecule Biophysics: RNA Splicing and DNA-Directed Enzyme Cascades

14. 7. 2014, 13.00 – 14.00 RCPTM, Šlechtitelů 27, Olomouc - Holice, místnost 314 (mapa)
Prof. Nils G. Walter

Abstract: Nature and Nanotechnology likewise employ nanoscale machines that self- assemble into structures of complex architecture and functionality. Fluorescence microscopy offers a non-invasive tool to probe and ultimately dissect and control these nanoassemblies in real-time. In particular, single molecule fluorescence resonance energy transfer (smFRET) allows us to measure distances at the 2-8 nm scale, whereas complementary super-resolution localization techniques based on Gaussian fitting of imagedpointspreadfunctions(PSFs)measuredistancesinthe10nmandlongerrange. Here,I will describe how we used smFRET to show that single spliceosomes responsible for the accurate removal of all intervening sequences (introns) in pre-messenger RNAs are working as biasedBrownianratchetmachines. Ontheotherendofthespectrum,wehaveutilizedsmFRET and super-resolution fluorescence microscopy to monitor enhanced enzyme cascades engineered to self-assemble on DNA origami. We can even "PAINT" these "nanopegboards", and watch them at super-resolution diffuse as DNA barges over supported lipid bilayers, one molecule/particle at a time.