title: Structural basis of transcriptional pausing in bacteria.
abstract: Transcriptional pausing by multisubunit RNA polymerases (RNAPs) is a key mechanism for regulating gene expression in both prokaryotes and eukaryotes and is a prerequisite for transcription termination. Pausing and termination states are thought to arise through a common, elemental pause state that is inhibitory for nucleotide addition. We report three crystal structures of Thermus RNAP elemental paused elongation complexes (ePECs). The structures reveal the same relaxed, open-clamp RNAP conformation in the ePEC that may arise by failure to re-establish DNA contacts during translocation. A kinked bridge-helix sterically blocks the RNAP active site, explaining how this conformation inhibits RNAP catalytic activity. Our results provide a framework for understanding how RNA hairpin formation stabilizes the paused state and how the ePEC intermediate facilitates termination.
authors: The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors: Albert
authors: A
authors: Weixlbaumer
authors: Katherine
authors: K
authors: Leon
authors: Robert
authors: R
authors: Landick
authors: Seth A
authors: SA
authors: Darst
mesh_terms: D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000378:metabolism; D055029:Inverted Repeat Sequences; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D013824:Thermus / Q000201:enzymology*; D016963:Thermus thermophilus / Q000201:enzymology*; D061805:Transcription Elongation, Genetic; D061806:Transcription Termination, Genetic
chemical_list: D009711:Nucleotides; D012333:RNA, Messenger; D012321:DNA-Directed RNA Polymerases
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