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Journal of Bacteriology, June 2004, p. 3848-3854, Vol. 186, No. 12
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.12.3848-3854.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Arginine Regulatory Protein Mediates Repression by Arginine of the Operons Encoding Glutamate Synthase and Anabolic Glutamate Dehydrogenase in Pseudomonas aeruginosa

Shehab Hashim,1 Dong-Hyeon Kwon,2 Ahmed Abdelal,3 and Chung-Dar Lu1*

Department of Biology, Georgia State University, Atlanta, Georgia 30303,1 Department of Biology, Northeastern University, Boston, Massachusetts 02115-5000,3 VA Medical Center, Baylor College of Medicine, Houston, Texas 770302

Received 10 December 2003/ Accepted 25 February 2004

The arginine regulatory protein of Pseudomonas aeruginosa, ArgR, is essential for induction of operons that encode enzymes of the arginine succinyltransferase (AST) pathway, which is the primary route for arginine utilization by this organism under aerobic conditions. ArgR also induces the operon that encodes a catabolic NAD+-dependent glutamate dehydrogenase (GDH), which converts L-glutamate, the product of the AST pathway, in {alpha}-ketoglutarate. The studies reported here show that ArgR also participates in the regulation of other enzymes of glutamate metabolism. Exogenous arginine repressed the specific activities of glutamate synthase (GltBD) and anabolic NADP-dependent GDH (GdhA) in cell extracts of strain PAO1, and this repression was abolished in an argR mutant. The promoter regions of the gltBD operon, which encodes GltBD, and the gdhA gene, which encodes GdhA, were identified by primer extension experiments. Measurements of ß-galactosidase expression from gltB::lacZ and gdhA::lacZ translational fusions confirmed the role of ArgR in mediating arginine repression. Gel retardation assays demonstrated the binding of homogeneous ArgR to DNA fragments carrying the regulatory regions for the gltBD and gdhA genes. DNase I footprinting experiments showed that ArgR protects DNA sequences in the control regions for these genes that are homologous to the consensus sequence of the ArgR binding site. In silica analysis of genomic information for P. fluorescens, P. putida, and P. stutzeri suggests that the findings reported here regarding ArgR regulation of operons that encode enzymes of glutamate biosynthesis in P. aeruginosa likely apply to other pseudomonads.

* Corresponding author. Mailing address: Department of Biology, Georgia State University, 24 Peachtree Center Ave., Atlanta, GA 30303. Phone: (404) 651-2531. Fax: (404) 651-2509. E-mail: biocdl{at}panther.gsu.edu.

Journal of Bacteriology, June 2004, p. 3848-3854, Vol. 186, No. 12
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.12.3848-3854.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Li, W., Lu, C.-D. (2007). Regulation of Carbon and Nitrogen Utilization by CbrAB and NtrBC Two-Component Systems in Pseudomonas aeruginosa. J. Bacteriol. 189: 5413-5420 [Abstract] [Full Text]  
  • Yang, Z., Lu, C.-D. (2007). Functional Genomics Enables Identification of Genes of the Arginine Transaminase Pathway in Pseudomonas aeruginosa. J. Bacteriol. 189: 3945-3953 [Abstract] [Full Text]  
  • Elbahloul, Y., Steinbuchel, A. (2006). Engineering the Genotype of Acinetobacter sp. Strain ADP1 To Enhance Biosynthesis of Cyanophycin. Appl. Environ. Microbiol. 72: 1410-1419 [Abstract] [Full Text]  
  • Lu, C.-D., Yang, Z., Li, W. (2004). Transcriptome Analysis of the ArgR Regulon in Pseudomonas aeruginosa. J. Bacteriol. 186: 3855-3861 [Abstract] [Full Text]  


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