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Journal of Bacteriology, September 2004, p. 6059-6069, Vol. 186, No. 18
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.18.6059-6069.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Two Arginine Repressors Regulate Arginine Biosynthesis in Lactobacillus plantarum

Hervé Nicoloff,1 Florence Arsène-Ploetze,1 Cédric Malandain,1 Michiel Kleerebezem,2 and Françoise Bringel1*

Laboratoire de Dynamique, Evolution et Expression de Génomes de Microorganismes, Université Louis Pasteur/CNRS FRE 2326, Strasbourg, France,1 Wageningen Centre for Food Sciences, Wageningen, The Netherlands2

Received 29 January 2004/ Accepted 9 June 2004

The repression of the carAB operon encoding carbamoyl phosphate synthase leads to Lactobacillus plantarum FB331 growth inhibition in the presence of arginine. This phenotype was used in a positive screening to select spontaneous mutants deregulated in the arginine biosynthesis pathway. Fourteen mutants were genetically characterized for constitutive arginine production. Mutations were located either in one of the arginine repressor genes (argR1 or argR2) present in L. plantarum or in a putative ARG operator in the intergenic region of the bipolar carAB-argCJBDF operons involved in arginine biosynthesis. Although the presence of two ArgR regulators is commonly found in gram-positive bacteria, only single arginine repressors have so far been well studied in Escherichia coli or Bacillus subtilis. In L. plantarum, arginine repression was abolished when ArgR1 or ArgR2 was mutated in the DNA binding domain, or in the oligomerization domain or when an A123D mutation occurred in ArgR1. A123, equivalent to the conserved residue A124 in E. coli ArgR involved in arginine binding, was different in the wild-type ArgR2. Thus, corepressor binding sites may be different in ArgR1 and ArgR2, which have only 35% identical residues. Other mutants harbored wild-type argR genes, and 20 mutants have lost their ability to grow in normal air without carbon dioxide enrichment; this revealed a link between arginine biosynthesis and a still-unknown CO2-dependent metabolic pathway. In many gram-positive bacteria, the expression and interaction of different ArgR-like proteins may imply a complex regulatory network in response to environmental stimuli.

* Corresponding author. Mailing address: Laboratoire de Dynamique, Evolution et Expression de Génomes de Microorganismes, Université Louis Pasteur/CNRS FRE 2326, 28 rue Goethe, 67083 Strasbourg, France. Phone: 33 3 90 24 18 15. Fax: 33 3 90 24 20 28. E-mail: bringel{at}gem.u-strasbg.fr.

Journal of Bacteriology, September 2004, p. 6059-6069, Vol. 186, No. 18
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.18.6059-6069.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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