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Journal of Bacteriology, March 2007, p. 1675-1688, Vol. 189, No. 5
0021-9193/07/$08.00+0     doi:10.1128/JB.00592-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Regulation of Sulfur Assimilation Pathways in Burkholderia cenocepacia: Identification of Transcription Factors CysB and SsuR and Their Role in Control of Target Genes{triangledown}

Roksana Iwanicka-Nowicka,1 Agata Zielak,1 Anne M. Cook,2 Mark S. Thomas,2 and Monika M. Hryniewicz1*

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland,1 Division of Genomic Medicine, School of Medicine and Biomedical Sciences, University of Sheffield, Sheffield, United Kingdom2

Received 26 April 2006/ Accepted 13 September 2006

Two genes encoding transcriptional regulators involved in sulfur assimilation pathways in Burkholderia cenocepacia strain 715j have been identified and characterized functionally. Knockout mutations in each of the B. cenocepacia genes were constructed and introduced into the genome of 715j by allelic replacement. Studies on the utilization of various sulfur sources by 715j and the obtained mutants demonstrated that one of the B. cenocepacia regulators, designated CysB, is preferentially involved in the control of sulfate transport and reduction, while the other, designated SsuR, is required for aliphatic sulfonate utilization. Using transcriptional promoter-lacZ fusions and DNA-binding experiments, we identified several target promoters for positive control by CysB and/or SsuR—sbpp (preceding the sbp cysT cysW cysA ssuR cluster), cysIp (preceding the cysI cysD1 cysN cysH cysG cluster), cysD2p (preceding a separate cluster, cysD2 cysNC), and ssuDp (located upstream of the ssuDCB operon)—and we demonstrated overlapping functions of CysB and SsuR at particular promoters. We also demonstrated that the cysB gene is negatively controlled by both CysB and SsuR but the ssuR gene itself is not significantly regulated as a separate transcription unit. The function of B. cenocepacia CysB (in vivo and in vitro) appeared to be independent of the presence of acetylserine, the indispensable coinducer of the CysB regulators of Escherichia coli and Salmonella. The phylogenetic relationships among members of the "CysB family" in the {gamma} and ß subphyla are presented.

* Corresponding author. Mailing address: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5A, 02-106 Warsaw, Poland. Phone: 48 22 592 13 06. Fax: 48 39 121 623. E-mail: monikah{at}ibb.waw.pl.

{triangledown} Published ahead of print on 22 September 2006.

Journal of Bacteriology, March 2007, p. 1675-1688, Vol. 189, No. 5
0021-9193/07/$08.00+0     doi:10.1128/JB.00592-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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