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Journal of Bacteriology, August 2005, p. 5648-5657, Vol. 187, No. 16
0021-9193/05/$08.00+0     doi:10.1128/JB.187.16.5648-5657.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

FpvIR Control of fpvA Ferric Pyoverdine Receptor Gene Expression in Pseudomonas aeruginosa: Demonstration of an Interaction between FpvI and FpvR and Identification of Mutations in Each Compromising This Interaction

Gyula Alan Rédly and Keith Poole*

Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada K7L 3N6

Received 29 March 2005/ Accepted 20 May 2005

FpvR is a presumed cytoplasmic membrane-associated anti-sigma factor that controls the activities of extracytoplasmic function sigma factors PvdS and FpvI responsible for transcription of pyoverdine biosynthetic genes and the ferric pyoverdine receptor gene, fpvA, respectively. Using deletion analysis and an in vivo bacterial two-hybrid system, FpvR interaction with these {sigma} factors was confirmed and shown to involve the cytoplasmic N-terminal 67 amino acid resides of FpvR. FpvR bound specifically to a C-terminal region of FpvI corresponding to region 4 of the {sigma}70 family of sigma factors. FpvR and FpvI mutant proteins compromised for this interaction were generated by random and site-directed PCR mutagenesis and invariably contained secondary structure-altering proline substitution in predicted {alpha}-helices within the FpvR N terminus or FpvI region 4. PvdS was shown to bind to the same N-terminal region of FpvR, and FpvR mutations compromising FpvI binding also compromised PvdS binding, although some mutations had a markedly greater impact on PvdS binding. Apparently, these two {sigma} factors bind to FpvR in a substantially similar but not identical fashion. Intriguingly, defects in FpvR binding correlated with a substantial drop in yields of the FpvI and to a lesser extent PvdS {sigma} factors, suggesting that FpvR-bound FpvI and PvdS are stable while free and active sigma factor is prone to turnover.


* Corresponding author. Mailing address: Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada K7L 3N6. Phone: (613) 533-6677. Fax: (613) 533-6796. E-mail: poolek{at}post.queensu.ca.


Journal of Bacteriology, August 2005, p. 5648-5657, Vol. 187, No. 16
0021-9193/05/$08.00+0     doi:10.1128/JB.187.16.5648-5657.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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