PMID: 9448277Mar 14, 1998Paper

In vivo genomic footprinting analysis reveals that the complex Bradyrhizobium japonicum fixRnifA promoter region is differently occupied by two distinct RNA polymerase holoenzymes

Proceedings of the National Academy of Sciences of the United States of America
H BarriosE Morett

Abstract

The Bradyrhizobium japonicum fixRnifA operon is transcribed from two promoters: fixRp1, a -24/-12 promoter recognized by the sigma54-holoenzyme form of the RNA polymerase, and fixRp2, a -35/-10 promoter that is transcribed by a second, unidentified, form of RNA polymerase holoenzyme. The fixRp1 promoter is autoregulated during microaerobiosis by NifA, whereas fixRp2 is also activated, but by a different regulatory protein. The main transcription start sites for these promoters are just two nucleotides apart, such that the conserved -12 and -10 regions of fixRp1 and fixRp2, respectively, must overlap each other, whereas the -24 and -35 regions lie one DNA helical turn apart. Using in vivo genomic dimethyl sulfate and KMnO4 footprinting, we showed that the promoter region is differentially protected, depending upon which holoenzyme is bound. Mutagenesis analyses indicated that positions from -12 to -14 are critical for the activity of both promoters, whereas mutations at -10 and -11 affected mainly fixRp2 expression. When the sequence of the putative -35 region of fixRp2 was modified to match the putative consensus, expression from this promoter was increased 3-fold and the reactivity toward KMnO4, but not the transcriptional sta...Continue Reading

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Citations

Oct 28, 1999·Nucleic Acids Research·H BarriosE Morett
Oct 24, 2007·Journal of Bacteriology·Andrea LindemannHans-Martin Fischer
Jan 27, 2011·Biochemical Society Transactions·María J TorresMaría J Delgado
Oct 10, 2003·Nucleosides, Nucleotides & Nucleic Acids·Chinh T BuiRichard G H Cotton
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·S E Rokita
Feb 11, 1999·Journal of Bacteriology·R S Janakiraman, Y V Brun

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