PMID: 9185571Jul 1, 1997Paper

sigma factor mutations affecting the sequence-selective interaction of RNA polymerase with -10 region single-stranded DNA

Nucleic Acids Research
X HuangJ D Helmann

Abstract

Thesigmasubunit of RNA polymerase determines promoter recognition and catalyzes DNA strand separation. The -35 promoter region is recognized by a helix-turn-helix motif in region 4, while the -10 region is specified, at least in part, by an amphipathic helix in region 2. We have proposed that conserved aromatic residues insigmaregion 2.3 interact with the non-template strand of the -10 element to drive open complex formation. We now report that Bacillus subtilis sigmaA holoenzyme, but neither core nor sigmaA alone, binds with high selectivity to single-stranded (ss) DNA containing the non-template -10 consensus sequence. UV irradiation of holoenzyme-ssDNA complexes efficiently crosslinks sigmaA to DNA and protease mapping supports a primary contact site in or near region 2. Several mutations in sigmaA region 2.3, shown previously to impair promoter melting, affect ssDNA binding: Y184A decreases binding selectivity, while Y189A and W193A decrease the efficiency of photocrosslinking. These results support a model in which these aromatic amino acids are juxtaposed to ssDNA, consistent with their demonstrated role in stabilizing the open complex.

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Citations

Apr 1, 2011·Journal of Biosciences·Olga N KorolevaValeriy L Drutsa
May 7, 2003·Current Opinion in Microbiology·Sergei Borukhov, Evgeny Nudler
May 10, 2002·Nature·Dmitry G VassylyevShigeyuki Yokoyama
Nov 13, 2001·Molecular Microbiology·N E Baldwin, A J Dombroski
Sep 6, 2011·Nucleosides, Nucleotides & Nucleic Acids·Olga Nikolaevna KorolevaValeriy Lvovich Drutsa
Oct 12, 2010·Nucleic Acids Research·Hsin-Yi YehBan-Yang Chang
Oct 17, 2009·Genes & Development·Andrey Feklistov, Seth A Darst
Jun 29, 1999·Cold Spring Harbor Symposia on Quantitative Biology·R H Ebright
Dec 2, 2010·Microbiology and Molecular Biology Reviews : MMBR·David BikardDidier Mazel
Nov 18, 2006·Journal of Zhejiang University. Science. B·Andrey Yu BerezhnoyKiryu Hisanori
Jul 29, 2016·Frontiers in Microbiology·Orlando SantillánGuillermo Dávila
Sep 20, 2005·The Journal of Biological Chemistry·Anita Niedziela-Majka, Tomasz Heyduk
Feb 26, 2005·The Journal of Biological Chemistry·Lisa A Schroeder, Pieter L deHaseth
Mar 20, 1999·The Journal of Biological Chemistry·O L Schaubach, A J Dombroski
Jun 11, 1998·Journal of Bacteriology·P L deHasethM T Record
Jan 27, 1999·Journal of Molecular Biology·M BuckleH Buc

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