Transcript analysis reveals an extended regulon and the importance of protein-protein co-operativity for the Escherichia coli methionine repressor

The Biochemical Journal
Ferenc MarincsP G Stockley

Abstract

We have used DNA arrays to investigate the effects of knocking out the methionine repressor gene, metJ, on the Escherichia coli transcriptome. We assayed the effects in the knockout strain of supplying wild-type or mutant MetJ repressors from an expression plasmid, thus establishing a rapid assay for in vivo effects of mutations characterized previously in vitro. Repression is largely restricted to known genes involved in the biosynthesis and uptake of methionine. However, we identified a number of additional genes that are significantly up-regulated in the absence of repressor. Sequence analysis of the 5' promoter regions of these genes identified plausible matches to met-box sequences for three of these, and subsequent electrophoretic mobility-shift assay analysis showed that for two such loci their repressor affinity is higher than or comparable with the known metB operator, suggesting that they are directly regulated. This can be rationalized for one of the loci, folE, by the metabolic role of its encoded enzyme; however, the links to the other regulated loci are unclear, suggesting both an extension to the known met regulon and additional complexity to the role of the repressor. The plasmid gene replacement system has been...Continue Reading

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Citations

Mar 18, 2009·Proceedings of the National Academy of Sciences of the United States of America·Anne M AugustusLeonard D Spicer
Nov 2, 2012·Molecular Plant-microbe Interactions : MPMI·Marion F CubittGeorge P C Salmond
Oct 22, 2008·Journal of Bacteriology·Gabriella PhillipsValérie de Crécy-Lagard
Nov 4, 2008·Journal of Bacteriology·Julio Collado-VidesAlberto Santos-Zavaleta
Nov 16, 2011·BMC Genomics·Anne M Augustus, Leonard D Spicer
Dec 5, 2006·PLoS Computational Biology·Rutger HermsenPieter Rein ten Wolde
Nov 26, 2009·PLoS Computational Biology·Wolfram StackliesFrauke Gräter
Apr 23, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jeremiah J TrauschRobert T Batey
Jun 14, 2014·Proceedings of the National Academy of Sciences of the United States of America·Rembrandt J F HaftRobert Landick
May 6, 2008·Journal of Molecular Biology·James A GarnettSimon E V Phillips
Dec 13, 2007·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Sep 12, 2006·The Journal of Biological Chemistry·Anne Marie AugustusLeonard D Spicer
Nov 16, 2016·Journal of Industrial Microbiology & Biotechnology·Hua LiGui Yang Shi

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