Abstract
In this communication, we present a genetic analysis of the glnN promoter region of Synechococcus elongatus PCC 7942. luxAB reporter fusions were used to characterize the glnN promoter by deletion and site-directed mutational analysis. Reporter gene expression analysis was performed in S. elongatus wild-type and mutant strains to reveal the role of the global nitrogen responsive transcription factor NtcA and of the PII signalling protein on regulation of glnN gene expression. A non-canonical NtcA-binding motif is responsible for NtcA-dependent, nitrogen-responsive regulation of glnN. The PII signalling protein has opposing effects on NtcA-dependent glnN expression. Under conditions of nitrate-growth, it depresses expression, whereas under conditions of combined nitrogen starvation, it is required for full induction. Furthermore, sequences upstream of the NtcA-binding site have repressive effect on glnN promoter activity.
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Acknowledgments
We are indebted to Susan S. Golden (Texas A&M University) for providing the reporter plasmid pAM1580 and to Peter Wolk (Michigan State University) for providing plasmids pRL542 and pRL443. We thank Peter Friedhoff and Laura Manelyte (Biochemistry, Giessen) for their help for generating the site-directed mutagenesis constructs and Katrin Lührsen for excellent DNA sequence analysis. M. Fadi Aldehni was a fellow from the Graduiertenkolleg “Biochemie der Nucleoproteinkomplexe.” This work was supported by a grant from the Deutsche Forschungsgemeinschaft (Fo 195/4).
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Aldehni, M.F., Forchhammer, K. Analysis of a non-canonical NtcA-dependent promoter in Synechococcus elongatus and its regulation by NtcA and PII . Arch Microbiol 184, 378–386 (2006). https://doi.org/10.1007/s00203-005-0056-6
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DOI: https://doi.org/10.1007/s00203-005-0056-6