Activation of the Rhizobium leguminosarum glnII gene by NtrC is dependent on upstream DNA sequences
Tóm tắt
The cloning and sequence determination is reported of the DNA region of Rhizobium leguminosarum coding for glutamine synthetase II (GSII). An open reading frame (ORF) encoding 326 amino acids was defined as the glnII gene on the basis of its similarity to other glnII genes and the ability of a DNA fragment carrying this ORF to complement the glutamine auxotrophy of a Klebsiella pneumoniae glnA mutant. We find that the glnII gene in R. leguminosarum is transcribed as a monocistronic unit from a single promoter, which shows structural features characteristic of rpoN(ntrA)-dependent promoters. In K. pneumoniae, such promoters require the ntrC and rpoN(ntrA) gene products for transcription. The intracellular level of glnII mRNA changes when R. leguminosarum is grown on different nitrogen sources, as expected for regulation by the nitrogen regulatory system. Promoter deletion analysis has shown that an extensive upstream DNA sequence (316 bp) is essential for in vivo activation of the glnII promoter in different biovars of R. leguminosarum. This DNA region requires a wild-type ntrC gene for activity and includes two conserved putative NtrC-binding site sequences. The results conclusively show that transcription from the R. leguminosarum glnII promoter is fully dependent on positive control by NtrC protein and on an upstream activator sequence (UAS).
Tài liệu tham khảo
Austin S, Henderson N, Dixon R (1987) Requirements for transcriptional activation in vitro of the nitrogen-regulated glnA and nifLA promoters from Klebsiella pneumoniae: dependence on activator concentration. Mol Microbiol 1:92–100
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Carlson TA, Chelm BK (1986) Apparent eukaryotic origin of glutamine synthetase II from the bacterium B. japonicum. Nature 321:568–570
Carlson TA, Martin GB, Chelm BK (1987) Differential transcription of the two glutamine synthetase genes of Bradyrhizobium japonicum. J Bacteriol 169:5861–5866
Chiurazzi M, Iaccarino M (1990) Transcriptional analysis of the glnB-glnA region of Rhizobium leguminosarum biovar viciae. Mol Microbiol 4:1727–1735
de Brujin FJ, Rossbach S, Schneider M, Radet P, Messmer S, Szeto WW, Ausubel FM, Schell J (1989) Rhizobium meliloti 1021 has three differentially regulated loci involved in glutamine biosynthesis, none of which is essential for symbiotic nitrogen fixation. J Bacteriol 171:1673–1682
Dixon R, Kennedy C, Kondorosi A, Krishnapillai V, Merrick M (1977) Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet 157:189–198
Drummond M, Clements J, Merrick M, Dixon R (1983) Positive control and autogenous regulation of nifLA promoter in Klebsiella penumoniae. Nature 301:302–307
Friedman AM, Long SR, Brown SE, Buikema WJ, Ausubel FM (1982) Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene 18:289–296
George DG, Baker WC, Hunt LT (1986) The protein identification resource (PIR). Nucleic Acids Res 14:11–16
Gussin GN, Ronson CW, Ausubel FM (1986) Regulation of nitrogen fixation genes. Annu Rev Genet 20:567–591
Hill RT, Parker JR, Goodman HJK, Jones DT, Woods DR (1989) Molecular analysis of a novel glutamine synthetase of the anaerobe Bacteroides fragilis. J Gen Microbiol 135:3271–3279
Holtel A, Merrick M (1988) Identification of the Klebsiella pneumoniae glnB gene: nucleotide sequence of wild-type and mutant alleles. Mol Gen Genet 215:134–138
Hooykaas PJJ, Clapwijk PM, Nuti MP, Shilperoort RA, Roersch A (1977) Transfer of the Agrobacterium tumefaciens T1 plasmid to avirulent Agrobacteria and to Rhizobium ex-planta. J Gen Microbiol 98:477–484
Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Martin GB, Chelm BK (1991) Bradyrhizobium japonicum ntrBC/glnA and nifA/glnA mutants: further evidence that separate regulatory pathways govern glnII expression in free-living and symbiotic cells. Mol Plant-Microbe Interact 4:254–261
Martin GB, Chapman KA, Chelm BK (1988) Role of the Bradyrhizobium japonicum ntrC gene product in differential regulation of the glutamine synthetase II gene (glnII). J Bacteriol 170:5452–5459
Martin GB, Thomashow MF, Chelm BK (1989) Bradyrhizobium japonicum glnB, and putative nitrogen-regulatory gene, is regulated by NtrC at tandem promoters. J Bacteriol 171:5638–5645
Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Moreno S, Patriarca EJ, Chiurazzi M, Meza R, Defez R, Iaccarino M, Espin G (1992) Phenotype of a Rhizobium leguminosarum ntrC mutant. Res Microbiol 143:161–171
Morett E, Buck M (1989) In vivo studies on the interaction of RNA polymerase sigma-54 with the Klebsiella pneumoniae and Rhizobium meliloti nifH promoters. J Mol Biol 21:65–77
Morett E, Moreno S, Espin G (1988) Transcription analysis of the three nifH genes of Rhizobium phaseoli with gene fusions. Mol Gen Genet 213:499–504
Noel KD, Sanchez A, Fernandez L, Leemans J, Cevallos MA (1984) Rhizobium phaseoli symbiotic mutants with transposon Tn5 interactions. J Bacteriol 158:148–155
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Popham DL, Szeto D, Keener J, Kustu S (1989) Functions of a bacterial activator protein that binds to transcriptional enhancers. Science 243:629–635
Reitzer LJ, Magasanik B (1985) Expression of glnA in Escherichia coli is regulated at tandem promoters. Proc Natl Acad Sci USA 82:1979–1983
Reitzer LJ, Magasanik B (1986) Transcription of glnA in E. coli is stimulated by activator bound to sites far from the promoter. Cell 45:785–792
Ronson CW, Nixon BT, Albright LM, Ausubel FM (1987) Rhizobium meliloti ntrA (rpoN) gene is required for diverse metabolic functions. J Bacteriol 169:2424–2431
Rossbach S, Schell J, de Bruijn FJ (1988) Cloning and analysis of Agrobacterium tumefaciens C58 loci involved in glutamine biosynthesis: neither the glnA (GSI) nor the glnII (GSII) gene plays a special role in virulence. Mol Gen Genet 212:38–47
Rossen L, Shearman LA, Johnston AWB, Downie JA (1985) The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant excudate induces the nodA, B, C genes. EMBO J 4:3369–3374
Rossi M, Defez R, Chiurazzi M, Lamberti A, Fuggi A, Iaccarino M (1989) Regulation of glutamine synthetase isoenzymes in Rhizobium leguminosarum biovar viciae. J Gen Microbiol 135:629–637
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Shatters RG, Kahn ML (1989) Glutamine synthetase II in Rhizobium: Reexamination of the proposed horizontal transfer of DNA from eukaryotes to prokaryotes. J Mol Evol 29:422–428.
Shatters RG, Somerville JE, Kahn ML (1989) Regulation of glutamine synthetase II activity in Rhizobium meliloti 104A14. J Bacteriol 171:5087–5094
Somerville JE, Shatters RG, Kahn ML (1989) Isolation, characterization and complementation of Rhizobium meliloti 104A14 mutants that lack glutamine synthetase II activity. J Bacteriol 171:5079–5086
Spaink HP, Okker RJH, Wijffelman CA, Pees E, Lugtenberg BJJ (1987) Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI. Plant Mol Biol 9:27–39
Sundaresan V, Ow DW, Ausubel FM (1983) Activation of Klebsiella pneumoniae and Rhizobium meliloti nitrogenase promoters by gln(ntr) regulatory proteins. Proc Natl Acad Sci USA 80:4030–4034
Szeto WW, Nixon BT, Ronson CW, Ausubel FM (1987) Identification and characterization of the Rhizobium meliloti ntrC gene: R. meliloti has separate regulatory pathways for an activation of nitrogen fixation genes in free-living and symbiotic cells. J Bacteriol 169:1423–1432
Thoeny B, Hennecke H (1989) The -24/-12 promoter comes of age. FEMS Microbiol Rev 63:341–358
Vingron M, Argos P (1989) A fast and sensitive multiple sequence alignment algorithm. CABIOS 5:115–121