Nitrogen availability and electron transport control the expression of glnB gene (encoding PII protein) in the cyanobacterium Synechocystis sp. PCC 6803.
The glnB gene from Synechocystis sp. PCC 6803 that encodes the PII protein has been cloned by heterologous hybridization using the corresponding glnB gene from Synechococcus sp. PCC 7942. An ORF of 336 nucleotides appeared that potentially coded for a protein of 112 amino acid residues (M(r) 12,397). The deduced amino acid sequence revealed a high identity (higher than 80%) with its cyanobacterial counterparts and a basal level of identity (close to 60%) with other PII proteins. A single mRNA of about 680 nucleotides was found under all growth conditions studied. glnB gene expression was specifically activated under nitrogen deprivation (a 10-fold increase respect to nitrogen-replete conditions). No differences in glnB mRNA levels were observed when using nitrate or ammonium as nitrogen sources. Amount of glnB mRNA decreased to undetectable levels when transferring cells to the dark, but effect was avoided by adding glucose to the culture medium. Primer extension analysis and band-shift assays indicated that expression of the glnB gene, elevated under nitrogen deprivation, might lie under the control of the nitrogen transcriptional regulator NtcA, although constitutive levels of expression were also detected from a sigma 70-dependent Escherichia coli-like promoter.
García-Domínguez
M
M
Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-CSIC, Centro de Investigaciones Científicas Isla de la Cartuja, Sevilla, Spain.
Florencio
F J
FJ
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003624:Darkness; D003991:Dibromothymoquinone / Q000494:pharmacology; D004237:Diuron / Q000494:pharmacology; D004579:Electron Transport / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005970:Glutamate Synthase / Q000235:genetics; D005974:Glutamate-Ammonia Ligase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008027:Light; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism* / Q000502:physiology; D051922:PII Nitrogen Regulatory Proteins; D045322:Photosynthetic Reaction Center Complex Proteins / Q000037:antagonists & inhibitors; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D051922:PII Nitrogen Regulatory Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D014157:Transcription Factors; C075617:ntcA protein, Synechococcus; D003991:Dibromothymoquinone; C404143:PIID regulatory protein, Bacteria; D004237:Diuron; D005970:Glutamate Synthase; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen