Correlation of Antagonistic Regulation ofleuOTranscription with the Cellular Levels of BglJ-RcsB and LeuO inEscherichia coli.
LeuO is a conserved and pleiotropic transcription regulator, antagonist of the nucleoid-associated silencer protein H-NS, and important for pathogenicity and multidrug resistance inEnterobacteriaceae. Regulation of transcription of theleuOgene is complex. It is silenced by H-NS and its paralog StpA, and it is autoregulated. In addition, inEscherichia coli leuOis antagonistically regulated by the heterodimeric transcription regulator BglJ-RcsB and by LeuO. BglJ-RcsB activatesleuO, while LeuO inhibits activation by BglJ-RcsB. Furthermore, LeuO activates expression ofbglJ, which is likewise H-NS repressed. Mutual activation ofleuOandbglJresembles a double-positive feedback network, which theoretically can result in bi-stability and heterogeneity, or be maintained in a stable OFF or ON states by an additional signal. Here we performed quantitative and single-cell expression analyses to address the antagonistic regulation and feedback control ofleuOtranscription by BglJ-RcsB and LeuO using aleuOpromotermVenusreporter fusion and finely tunablebglJandleuOexpression plasmids. The data revealed uniform regulation ofleuOexpression in the population that correlates with the relative cellular concentration of BglJ and LeuO. The data are in agreement with a straightforward model of antagonistic regulation ofleuOexpression by the two regulators, LeuO and BglJ-RcsB, by independent mechanisms. Further, the data suggest that at standard laboratory growth conditions feedback regulation ofleuOis of minor relevance and that silencing ofleuOandbglJby H-NS (and StpA) keeps these loci in the OFF state.
Department of Biology, Institute for Genetics, University of Cologne Cologne, Germany.
Hannes
H
Breddermann
Department of Biology, Institute for Genetics, University of Cologne Cologne, Germany.
Karin
K
Schnetz
D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D059010:Single-Cell Analysis; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000378:metabolism*; D014158:Transcription, Genetic
C103411:BglJ protein, E coli; D029968:Escherichia coli Proteins; C430340:LeuO protein, E coli; C489568:RcsB protein, E coli; D015534:Trans-Activators; D014157:Transcription Factors
H-NS; H-NS antagonist; feedback regulation; nucleoid-associated protein; transcription regulator
Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8168-72
9223333
J Bacteriol. 2010 Dec;192(24):6456-64
20952573
J Biol Chem. 2001 Jun 1;276(22):19046-51
11376008
J Bacteriol. 2014 Jun;196(12):2143-54
24659766
Proc Natl Acad Sci U S A. 1988 Dec;85(23):8973-7
3057497
Microbiology. 2008 Sep;154(Pt 9):2533-45
18757787
J Bacteriol. 1998 Feb;180(3):626-33
9457867
Biophys J. 2008 Aug;95(4):2103-15
18469087
J Biol Chem. 2005 Apr 15;280(15):15111-21
15711009
Mol Microbiol. 2010 Mar;75(6):1495-512
20132443
Curr Opin Microbiol. 2015 Apr;24:53-9
25638302
Nat Rev Genet. 2007 Jun;8(6):450-61
17510665
Mol Microbiol. 2012 Mar;83(6):1109-23
22295907
J Biol Chem. 2001 Mar 23;276(12):9478-85
11121424
Nat Rev Microbiol. 2010 Mar;8(3):185-95
20140026
Gene. 1995 May 26;158(1):9-14
7789817
J Bacteriol. 2011 May;193(10):2396-407
21398529
Cell. 2010 Oct 15;143(2):326-e1
20946989
J Bacteriol. 1989 Sep;171(9):4617-22
2548993
Mol Microbiol. 2010 Sep;77(6):1380-93
20659289
J Bacteriol. 2009 Jul;191(14):4562-71
19429622
Mol Microbiol. 2002 Jan;43(1):217-26
11849549
Genetics. 1996 Jun;143(2):627-35
8725214
Microbiology. 2008 Dec;154(Pt 12):3609-23
19047729
Annu Rev Microbiol. 2005;59:379-405
16153174
Mol Microbiol. 2007 Nov;66(3):727-43
17908208
J Biol Chem. 1989 Oct 15;264(29):16973-6
2676996
Mol Microbiol. 2012 Sep;85(6):1072-89
22804842
Nature. 1979 Jul 5;280(5717):80-2
15305587
J Biol Eng. 2014 Jan 02;8(1):2
24382032
J Biol Chem. 2003 Sep 26;278(39):38094-103
12871947
Mol Microbiol. 2011 Oct;82(2):378-97
21883529
Microbiology. 2001 Dec;147(Pt 12):3241-7
11739756
J Bacteriol. 1995 Jul;177(14):4121-30
7608087
Proc Natl Acad Sci U S A. 2004 Feb 17;101(7):1822-7
14766974
Biochem Biophys Res Commun. 2000 Sep 16;276(1):64-70
11006083
Nucleic Acids Res. 2014 Mar;42(5):2999-3008
24335284
Nucleic Acids Res. 2016 Mar 18;44(5):2058-74
26843427
Biophys J. 2015 Oct 6;109 (7):1321-9
26445432
Mol Microbiol. 1997 Aug;25(3):559-69
9302018
Eur J Biochem. 1993 Jan 15;211(1-2):181-91
8425528
J Bacteriol. 1998 Jan;180(1):190-3
9422614
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5
10829079
Gene. 1988 Sep 30;69(2):301-15
3069586
J Biol Chem. 2016 Jan 29;291(5):2357-70
26635367
J Bacteriol. 2008 Feb;190(3):926-35
18055596