Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Oct;177(20):5812–5817. doi: 10.1128/jb.177.20.5812-5817.1995

Phosphorylation of the PII protein (glnB gene product) in the cyanobacterium Synechococcus sp. strain PCC 7942: analysis of in vitro kinase activity.

K Forchhammer 1, N Tandeau de Marsac 1
PMCID: PMC177403  PMID: 7592328

Abstract

The PII protein in the cyanobacterium Synechococcus sp. strain PCC 7942 signals the cellular state of nitrogen assimilation relative to CO2 fixation by being phosphorylated at a seryl residue. In this study, we first determined the location of the phosphorylated seryl residue within the PII amino acid sequence. The phosphorylation site exhibits an RXS motif, a recognition sequence characteristic for cyclic AMP-dependent protein serine kinases from eukaryotes. We established an in vitro PII phosphorylation assay to further analyze the PII kinase activity in Synechococcus sp. strain PCC 7942. ATP was used specifically as a phosphoryl donor, and the PII kinase activity was shown to be stimulated by alpha-ketoglutarate. Unlike the PII-modifying uridylyltransferase- and uridylyl-removing enzyme characterized in proteobacteria, the activity of the PII kinase from the cyanobacterium did not respond to glutamine.

Full Text

The Full Text of this article is available as a PDF (352.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adler S. P., Purich D., Stadtman E. R. Cascade control of Escherichia coli glutamine synthetase. Properties of the PII regulatory protein and the uridylyltransferase-uridylyl-removing enzyme. J Biol Chem. 1975 Aug 25;250(16):6264–6272. [PubMed] [Google Scholar]
  2. Aiba H., Nagaya M., Mizuno T. Sensor and regulator proteins from the cyanobacterium Synechococcus species PCC7942 that belong to the bacterial signal-transduction protein families: implication in the adaptive response to phosphate limitation. Mol Microbiol. 1993 Apr;8(1):81–91. doi: 10.1111/j.1365-2958.1993.tb01205.x. [DOI] [PubMed] [Google Scholar]
  3. Atkinson M. R., Kamberov E. S., Weiss R. L., Ninfa A. J. Reversible uridylylation of the Escherichia coli PII signal transduction protein regulates its ability to stimulate the dephosphorylation of the transcription factor nitrogen regulator I (NRI or NtrC). J Biol Chem. 1994 Nov 11;269(45):28288–28293. [PubMed] [Google Scholar]
  4. Forchhammer K., Tandeau de Marsac N. Functional analysis of the phosphoprotein PII (glnB gene product) in the cyanobacterium Synechococcus sp. strain PCC 7942. J Bacteriol. 1995 Apr;177(8):2033–2040. doi: 10.1128/jb.177.8.2033-2040.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Forchhammer K., Tandeau de Marsac N. The PII protein in the cyanobacterium Synechococcus sp. strain PCC 7942 is modified by serine phosphorylation and signals the cellular N-status. J Bacteriol. 1994 Jan;176(1):84–91. doi: 10.1128/jb.176.1.84-91.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Herrero A., Flores E., Guerrero M. G. Regulation of nitrate reductase levels in the cyanobacteria Anacystis nidulans, Anabaena sp. strain 7119, and Nostoc sp. strain 6719. J Bacteriol. 1981 Jan;145(1):175–180. doi: 10.1128/jb.145.1.175-180.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kemp B. E., Pearson R. B. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. doi: 10.1016/0968-0004(90)90073-k. [DOI] [PubMed] [Google Scholar]
  8. Kuhlemeier C. J., Thomas A. A., van der Ende A., van Leen R. W., Borrias W. E., van den Hondel C. A., van Arkel G. A. A host-vector system for gene cloning in the cyanobacterium Anacystis nidulans R2. Plasmid. 1983 Sep;10(2):156–163. doi: 10.1016/0147-619x(83)90068-9. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Liang J., Scappino L., Haselkorn R. The patA gene product, which contains a region similar to CheY of Escherichia coli, controls heterocyst pattern formation in the cyanobacterium Anabaena 7120. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5655–5659. doi: 10.1073/pnas.89.12.5655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mangum J. H., Magni G., Stadtman E. R. Regulation of glutamine synthetase adenylylation and deadenylylation by the enzymatic uridylylation and deuridylylation of the PII regulatory protein. Arch Biochem Biophys. 1973 Oct;158(2):514–525. doi: 10.1016/0003-9861(73)90543-2. [DOI] [PubMed] [Google Scholar]
  12. Mann N. H. Protein phosphorylation in cyanobacteria. Microbiology. 1994 Dec;140(Pt 12):3207–3215. doi: 10.1099/13500872-140-12-3207. [DOI] [PubMed] [Google Scholar]
  13. Ninfa A. J., Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909–5913. doi: 10.1073/pnas.83.16.5909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reyes J. C., Florencio F. J. A mutant lacking the glutamine synthetase gene (glnA) is impaired in the regulation of the nitrate assimilation system in the cyanobacterium Synechocystis sp. strain PCC 6803. J Bacteriol. 1994 Dec;176(24):7516–7523. doi: 10.1128/jb.176.24.7516-7523.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rosenfeld J., Capdevielle J., Guillemot J. C., Ferrara P. In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis. Anal Biochem. 1992 May 15;203(1):173–179. doi: 10.1016/0003-2697(92)90061-b. [DOI] [PubMed] [Google Scholar]
  16. Stanier R. Y., Cohen-Bazire G. Phototrophic prokaryotes: the cyanobacteria. Annu Rev Microbiol. 1977;31:225–274. doi: 10.1146/annurev.mi.31.100177.001301. [DOI] [PubMed] [Google Scholar]
  17. Tsinoremas N. F., Castets A. M., Harrison M. A., Allen J. F., Tandeau de Marsac N. Photosynthetic electron transport controls nitrogen assimilation in cyanobacteria by means of posttranslational modification of the glnB gene product. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4565–4569. doi: 10.1073/pnas.88.11.4565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Zhang C. C. A gene encoding a protein related to eukaryotic protein kinases from the filamentous heterocystous cyanobacterium Anabaena PCC 7120. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11840–11844. doi: 10.1073/pnas.90.24.11840. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES