title: Single-molecule tracking of mRNA exiting from RNA polymerase II.
abstract: Single-pair fluorescence resonance energy transfer was used to track RNA exiting from RNA polymerase II (Pol II) in elongation complexes. Measuring the distance between the RNA 5' end and three known locations within the elongation complex allows us 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 nt, its 5' end associates with Pol II at the base of the dock domain. Because the initiation factor TFIIB binds to the dock domain and exit tunnel, exiting RNA may prevent TFIIB reassociation during elongation. RNA further extends toward the linker connecting to the polymerase C-terminal repeat domain (CTD), which binds the 5'-capping enzyme and other RNA processing factors.
authors.affiliation: Munich Center for Integrated Protein Science (CiPS) and Center for Nanoscience (CeNS), Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
authors.forename: Joanna
authors.initials: J
authors.lastname: Andrecka
authors.forename: Robert
authors.initials: R
authors.lastname: Lewis
authors.forename: Florian
authors.initials: F
authors.lastname: Brückner
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Lehmann
authors.forename: Patrick
authors.initials: P
authors.lastname: Cramer
authors.forename: Jens
authors.initials: J
authors.lastname: Michaelis
mesh_terms: D003545:Cysteine / Q000737:chemistry; D004257:DNA Polymerase II / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005656:Fungal Proteins / Q000737:chemistry*; D007834:Lasers; D008958:Models, Molecular; D015233:Models, Statistical; D008968:Molecular Conformation; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry; D012319:RNA Polymerase II / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry*; D035581:Transcription Factor TFIIB / Q000737:chemistry
chemical_list: D005656:Fungal Proteins; D009841:Oligonucleotides; D012333:RNA, Messenger; D035581:Transcription Factor TFIIB; D012313:RNA; D012319:RNA Polymerase II; D004257:DNA Polymerase II; D003545:Cysteine