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Journal of Bacteriology, March 2002, p. 1530-1539, Vol. 184, No. 6
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.6.1530-1539.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Bordetella Interspecies Allelic Variation in AlcR Inducer Requirements: Identification of a Critical Determinant of AlcR Inducer Responsiveness and Construction of an alcR(Con) Mutant Allele

Timothy J. Brickman* and Sandra K. Armstrong

Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota 55455-0312

Received 15 October 2001/ Accepted 13 December 2001

Previous studies established the critical roles of AlcR and alcaligin inducer in positive regulation of alcaligin siderophore biosynthesis and transport genes in Bordetella pertussis and Bordetella bronchiseptica. Transcriptional analyses using plasmid-borne alcR genes of B. pertussis UT25 and B. bronchiseptica B013N to complement the alcR defect of B. bronchiseptica strain BRM13 ({Delta}alcR1 alcA::mini-Tn5 lacZ1) revealed interspecies differences in AlcR inducer requirements for activation of alcABCDER operon transcription. Whereas the B. pertussis UT25 AlcR protein retained strong inducer dependence when produced from multicopy plasmids, B. bronchiseptica B013N alcR partially suppressed the alcaligin requirement for transcriptional activation. Functional analysis of AlcR chimeras produced by interspecies domain swapping and interspecies reciprocal site-specific mutagenesis determined that the phenotypic difference in AlcR inducer dependence was due to a single amino acid difference within the proposed inducer-binding and multimerization domain of AlcR. Structural predictions guided the design of a mutant AlcR protein with a single amino acid substitution at this critical position, AlcR(S103T), that was fully constitutive not only when produced from multicopy plasmids but also at a single-copy gene dosage. These results indicate that AlcR residue 103 affects a critical determinant of alcaligin inducer dependence of AlcR-mediated transcriptional activation. The alcR(S103T) mutant allele is the first alcR(Con) mutant allele identified.

* Corresponding author. Mailing address: Department of Microbiology, University of Minnesota, MMC 196 FUMC, 420 Delaware St. S.E., Minneapolis, MN 55455-0312. Phone: (612) 624-5177. Fax: (612) 626-0623. E-mail: brickman{at}lenti.med.umn.edu.

Journal of Bacteriology, March 2002, p. 1530-1539, Vol. 184, No. 6
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.6.1530-1539.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Brickman, T. J., Armstrong, S. K. (2005). Bordetella AlcS Transporter Functions in Alcaligin Siderophore Export and Is Central to Inducer Sensing in Positive Regulation of Alcaligin System Gene Expression. J. Bacteriol. 187: 3650-3661 [Abstract] [Full Text]  
  • Anderson, M. T., Armstrong, S. K. (2004). The BfeR Regulator Mediates Enterobactin-Inducible Expression of Bordetella Enterobactin Utilization Genes. J. Bacteriol. 186: 7302-7311 [Abstract] [Full Text]  
  • Vanderpool, C. K., Armstrong, S. K. (2003). Heme-Responsive Transcriptional Activation of Bordetella bhu Genes. J. Bacteriol. 185: 909-917 [Abstract] [Full Text]  


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