A new type of glutamine synthetase in cyanobacteria: the protein encoded by the glnN gene supports nitrogen assimilation in Synechocystis sp. strain PCC 6803.
A new glutamine synthetase gene, glnN, which encodes a polypeptide of 724 amino acid residues (M(r), 79,416), has been identified in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803; this is the second gene that encodes a glutamine synthetase (GS) in this cyanobacterium. The functionality of this gene was evidenced by its ability to complement an Escherichia coli glnA mutant and to support Synechocystis growth in a strain whose glnA gene was inactivated by insertional mutagenesis. In this mutant (strain SJCR3), as well as in the wild-type strain, the second GS activity was subject to regulation by the nitrogen source, being strongly enhanced in nitrogen-free medium. Transcriptional fusion of a chloramphenicol acetyltransferase (cat) gene with the 5'-upstream region of glnN suggested that synthesis of the second Synechocystis GS is regulated at the transcriptional level. Furthermore, the level of glnN mRNA, a transcript of about 2,300 bases, was found to be strongly increased in nitrogen-free medium. The glnN product is similar to the GS subunits of Bacteroides fragilis and Butyrivibrio fibrisolvens, two obligate anaerobic bacteria whose GSs are markedly different from other prokaryotic and eukaryotic GSs. However, significant similarity is evident in the five regions which are homologous in all of the GSs so far described. The new GS gene was also found in other cyanobacteria but not in N2-fixing filamentous species.
Reyes
J C
JC
Departamento de Bioquímica Vegetal y Biología Molecular, Universidad de Sevilla-CSIC, Spain.
Florencio
F J
FJ
D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology; D001483:Base Sequence; D000458:Cyanobacteria / Q000201:enzymology* / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D005974:Glutamate-Ammonia Ligase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism*; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen