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Genetics, Vol. 178, 185-195, January 2008, Copyright © 2008
doi:10.1534/genetics.107.081984

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Dual Involvement of CbrAB and NtrBC in the Regulation of Histidine Utilization in Pseudomonas fluorescens SBW25

Xue-Xian Zhang1 and Paul B. Rainey

Institute of Molecular Biosciences and New Zealand Institute for Advanced Study, Massey University, Auckland, New Zealand

1 Corresponding author: Institute of Molecular Biosciences, Massey University, Private Bag 102 904, North Shore Mail Centre, Auckland, New Zealand.
E-mail: x.x.zhang1{at}massey.ac.nz

Pseudomonas fluorescens SBW25 is capable of growing on histidine as a sole source of carbon and/or nitrogen. Previous work showed that the two-component regulatory system CbrAB is required for expression of the histidine utilization (hut) locus when histidine is the sole source of carbon and nitrogen. Here, using mutational analysis and transcriptional assays, we demonstrate involvement of a second two-component system, NtrBC. When histidine is the sole carbon source, transcription of the hutU operon is initiated from a {sigma}54-type promoter and requires CbrB (an enhancer binding protein for {sigma}54-recruitment). However, when histidine is the sole nitrogen source, the hutU operon is transcribed from a {sigma}70-type promoter and requires either CbrB or the nitrogen regulator, NtrC. No role was found for the SBW25 homolog of the nitrogen assimilation control protein (NAC). Biolog phenotypic microarray analysis of the ability of the three mutants ({Delta}cbrB, {Delta}ntrC, and {Delta}cbrB {Delta}ntrC) to utilize 190 carbon and 95 nitrogen substrates confirmed the central regulatory roles of CbrAB and NtrBC in cellular carbon and nitrogen catabolism: deletion of cbrB abolished growth on 20 carbon substrates; deletion of ntrC eliminated growth on 28 nitrogen substrates. A double cbrB–ntrC mutant was unable to utilize a further 14 nitrogen substrates (including histidine, proline, leucine, isoleucine, and valine). Our data show that CbrAB plays a role in regulation of both carbon and nitrogen catabolism and maintains activity of catabolic pathways under different C:N ratios.






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Copyright © 2008 by the Genetics Society of America.