Berlin 2008 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 26: Posters II
BP 26.34: Poster
Donnerstag, 28. Februar 2008, 17:00–19:30, Poster A
Single Molecule FRET studies of RNA Polymerese II — •Joanna Andrecka1, Robert Lewis1, Florian Brueckner2, Patrick Cramer2, and Jens Michaelis1 — 1Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstr.11, 81377 Munich, Germany — 2Gene Center Munich and Department of Chemistry and Biochemistry, Feodor-Lynen-Strasse 25, Ludwig-Maximilians-Universität München, 81377 Munich, Germany
Single-pair Fluorescence Resonance Energy Transfer (FRET) was used to track RNA exiting from RNA Polymerase (Pol II) in elongation complexes [1]. Measuring the distance between the RNA 5'-end and three known locations within the elongation complex allowed us to determine its position by means of triangulation. RNA leaves the polymerase active center cleft via the previously proposed exit tunnel, and then disengages from the enzyme surface. When the RNA reaches lengths of 26 and 29 nucleotides, its 5'-end associates with Pol II at the base of the dock domain. Since the initiation factor TFIIB binds to the dock domain and exit tunnel, exiting RNA may contribute to TFIIB displacement during the initiation to elongation transition and may prevent TFIIB re-association during elongation.
[1] J. Andrecka, R. Lewis, F. Brueckner, E. Lehmann, P. Cramer, J. Michaelis: Single-molecule tracking of mRNA exiting from RNA polymerase II, PNAS (accepted)