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Journal of Bacteriology, May 2008, p. 3374-3380, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.02002-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Evidence of In Vivo Cross Talk between the Nitrogen-Related and Fructose-Related Branches of the Carbohydrate Phosphotransferase System of Pseudomonas putida{triangledown}

Katharina Pflüger and Víctor de Lorenzo*

Centro Nacional de Biotecnología-CSIC, Campus de Cantoblanco, Madrid 28049, Spain

Received 24 December 2007/ Accepted 12 February 2008

The genome of Pseudomonas putida KT2440 encodes only five recognizable proteins belonging to the phosphoenolpyruvate (PEP)-carbohydrate phosphotransferase system (PTS). Two of these PTS constituents (FruA and FruB) form a complete system for fructose intake. The other three products, encoded by ptsP (EINtr), ptsO (NPr), and ptsN (EIIANtr), comprise a branch of the system unrelated to sugar traffic but thought to have an influence on coordination of N and C metabolism. We used a genetic approach to clarify the course of high-energy phosphate through this reduced set of PTS proteins. To this end, we monitored the phosphorylation state in vivo of the EIIANtr enzyme in various genetic backgrounds and growth conditions. Our results show that the source of phosphate available to the system is PEP and that the primary flow of phosphate through the N/C-sensing PTS proceeds from PEP to EINtr to NPr to EIIANtr. We also found that in the presence of fructose, unlike in the presence of succinate, EIIANtr can be phosphorylated in a ptsP strain but not in a ptsP fruB double mutant. This result revealed that the fructose transport system has the ability to cross talk in vivo with the N-related PTS branch. The data reported here thus document an unexpected connection in vivo between the sugar-dependent and sugar-independent PTSs.

* Corresponding author. Mailing address: Centro Nacional de Biotecnología-CSIC, Campus UAM-Cantoblanco, Madrid 28049, Spain. Phone: 34-91 585 45 36. Fax: 34-91 585 45 06. E-mail: vdlorenzo{at}cnb.csic.es

{triangledown} Published ahead of print on 22 February 2008.

Journal of Bacteriology, May 2008, p. 3374-3380, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.02002-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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