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Journal of Bacteriology, April 2003, p. 2582-2591, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2582-2591.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Signal Transduction Protein PII Is Required for NtcA-Regulated Gene Expression during Nitrogen Deprivation in the Cyanobacterium Synechococcus elongatus Strain PCC 7942

M. Fadi Aldehni,1 Jörg Sauer,1 Christian Spielhaupter,2,{dagger} Roland Schmid,3 and Karl Forchhammer1*

Institut für Mikrobiologie und Molekularbiologie der Justus-Liebig-Universität Giessen, D-35392 Giessen,1 Lehrstuhl für Mikrobiologie der Universität München, D-80638 München,2 Abteilung für Mikrobiologie der Universität Osnabrück, D-49069 Osnabrück, Germany3

Received 8 August 2002/ Accepted 17 December 2002

The transcription factor of the cyclic AMP receptor protein/FNR family, NtcA, and the PII signaling protein play central roles in global nitrogen control in cyanobacteria. A dependence on PII for NtcA-regulated transcription, however, has not been observed. In the present investigation, we examined alterations in gene expression following nitrogen deprivation in Synechococcus elongatus strain PCC 7942 and specifically the roles of NtcA and PII. Global changes in de novo protein synthesis following combined-nitrogen deprivation were visualized by in vivo [35S]methionine labeling and two-dimensional polyacrylamide gel electrophoresis analysis. Nearly all proteins whose synthesis responded specifically to combined-nitrogen deprivation in wild-type cells of S. elongatus failed to respond in PII- and NtcA-deficient mutants. One of the proteins whose synthesis was down-regulated in a PII- and NtcA-dependent manner was RbcS, the small subunit of RubisCO. Quantification of its mRNA revealed that the abundance of the rbcLS transcript following combined-nitrogen deprivation rapidly declined in wild-type cells but not in PII and NtcA mutant cells. To investigate further the relationship between PII and NtcA, fusions of the promotorless luxAB reporter genes to the NtcA-regulated glnB gene were constructed and these constructs were used to transform wild-type cells and PII- and NtcA- mutants. Determination of bioluminescence under different growth conditions showed that NtcA represses gene expression in the presence of ammonium in a PII-independent manner. By contrast, NtcA-dependent activation of glnB expression following combined-nitrogen deprivation was impaired in the absence of PII. Together, these results suggest that under conditions of combined-nitrogen deprivation, the regulation of NtcA-dependent gene expression requires the PII signal transduction protein.

* Corresponding author. Mailing address: Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany. Phone: 49 641 9935545. Fax: 49 641 9935549. E-mail: Karl.Forchhammer{at}mikro.bio.uni-giessen.de.

{dagger} Present address: Max von Pettenkofer-Institut für Virologie, Genzentrum München, D-81377 München, Germany.

Journal of Bacteriology, April 2003, p. 2582-2591, Vol. 185, No. 8
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.8.2582-2591.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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