Abstract
The nitrogen-fixing, root-associated strain Pseudomonas stutzeri A1501 carries a single gene encoding a protein from the PII family, designated glnK. The glnK gene is co-transcribed with two distantly related copies of amtB genes encoding putative ammonium channels. Transcription of glnK was decreased in the presence of ammonia and was partly dependent on NtrC and RpoN under nitrogen-limiting conditions. Inactivation of glnK led to a mutant strain devoid of nitrogenase activity, auxotrophic for glutamine and unable to deadenylylate glutamine synthetase, while inactivation of amtB 1 led to a prototrophic and Nif+ mutant strain. RT-PCR analysis showed that nifA transcription was abolished in the glnK mutant, while glnA remained transcribed. Using the yeast two-hybrid system, an interaction between GlnK and the C-terminal domain of NifL was observed, suggesting GlnK-dependent control of NifA activity by NifL. Introduction of a plasmid that expressed nifA from a constitutive promoter restored nitrogen fixation to the glnK mutant, and nitrogenase activity was observed even in the presence of ammonia. GlnK signalling appears to be a key regulatory element in control of ammonia assimilation, of nifA expression and in modulation of NifA activity by NifL.
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Acknowledgments
The authors wish to thank Mr. Dong-Jie Tang for suggesting to use the primer P18conF and Ms. Jerri Bram for reading the typescript. This work was supported by the Ministry of Science and Technology of China (National Basic Research Program 2001CB108904 and 2007CB707805, National High-Tech Program 2004AA21470 and 2006AA020101), and the National Natural Science Foundation of China (grant No. 30470047 and 30670050). Sheng He thanks College of Life Science and Technology of Guangxi University for support.
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Communicated by Jack Meeks.
S. He and M. Chen contributed equally to the work.
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He, S., Chen, M., Xie, Z. et al. Involvement of GlnK, a PII protein, in control of nitrogen fixation and ammonia assimilation in Pseudomonas stutzeri A1501. Arch Microbiol 190, 1–10 (2008). https://doi.org/10.1007/s00203-008-0354-x
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DOI: https://doi.org/10.1007/s00203-008-0354-x