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J Bacteriol. 2004 May;186(10):2921-7.

TtgV bound to a complex operator site represses transcription of the promoter for the multidrug and solvent extrusion TtgGHI pump.

Guazzaroni ME, Terán W, Zhang X, Gallegos MT, Ramos JL.

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Department of Biochemistry and Molecular and Cell Biology of Plants, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, E-18008 Granada, Spain.

Abstract

The TtgGHI efflux pump of Pseudomonas putida extrudes a variety of antibiotics and solvents. We show that the ttgGHI operon is transcribed in vitro and in vivo from a single promoter and not from two overlapping promoters as previously proposed. The expression of this promoter is controlled by the TtgV repressor, whose operator expands through four helical turns that overlap the -10 region of the promoter. We also show that TtgV is released from its operator on binding of effectors such as aliphatic alcohols. Mutational analysis of the ttgGHI promoter revealed that substitutions at -13, -12, and -8 yielded promoters that were unable to drive transcription whereas certain mutations at -9, -11, and -6 to -3 increased expression in vivo. The cause of the increased expression was either a decrease in the affinity of the TtgV protein for its operator or an increase in the affinity of RNA polymerase for the mutant promoters.

PMID:: 15126451; [PubMed - indexed for MEDLINE]
PMCID:: PMC400617

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Related citations in PubMed

In vivo and in vitro evidence that TtgV is the specific regulator of the TtgGHI multidrug and solvent efflux pump of Pseudomonas putida. [J Bacteriol. 2003]
In vivo and in vitro evidence that TtgV is the specific regulator of the TtgGHI multidrug and solvent efflux pump of Pseudomonas putida.
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Cited by 9 PubMed Central articles

Intramolecular signal transmission in a tetrameric repressor of the IclR family. [Proc Natl Acad Sci U S A. 2011]
Intramolecular signal transmission in a tetrameric repressor of the IclR family.
Fillet S, Krell T, Morel B, Lu D, Zhang X, Ramos JL. Proc Natl Acad Sci U S A. 2011 Sep 13; 108(37):15372-7. Epub 2011 Aug 29.
An antirepressor, SrpR, is involved in transcriptional regulation of the SrpABC solvent tolerance efflux pump of Pseudomonas putida S12. [J Bacteriol. 2011]
An antirepressor, SrpR, is involved in transcriptional regulation of the SrpABC solvent tolerance efflux pump of Pseudomonas putida S12.
Sun X, Zahir Z, Lynch KH, Dennis JJ. J Bacteriol. 2011 Jun; 193(11):2717-25. Epub 2011 Mar 25.
Crystal structure of TtgV in complex with its DNA operator reveals a general model for cooperative DNA binding of tetrameric gene regulators. [Genes Dev. 2010]
Crystal structure of TtgV in complex with its DNA operator reveals a general model for cooperative DNA binding of tetrameric gene regulators.
Lu D, Fillet S, Meng C, Alguel Y, Kloppsteck P, Bergeron J, Krell T, Gallegos MT, Ramos J, Zhang X. Genes Dev. 2010 Nov 15; 24(22):2556-65.

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