Localization of denitrification genes on the chromosomal map of Pseudomonas aeruginosa

Microbiology (United Kingdom) - Tập 144 Số 2 - Trang 441-448 - 1998
Kai‐Uwe Vollack1, Jieshuo Xie1, Elisabeth Härtig1, Ute Römling2, Walter G. Zumft1
1Lehrstuhl für Mikrobiologie, Universität Fridericiana, D-76128 Karlsruhe, Germany
2Karolinska Institutet, MTC, Department of Bacteriology, Box 280, Stockholm, Sweden

Tóm tắt

Cleavage of chromosomal DNA from Pseudomonas aeruginosa PAO by SpeI and DpnI has been used together with PFGE and Southern hybridization to establish the map location of the following principal denitrification genes: narGH (encoding the large and small subunits of respiratory nitrate reductase), nirS (cytochrome-cd 1 nitrite reductase), nirE (uroporphyrinogen-III methyltransferase for haem d 1 biosynthesis), norCB (nitric-oxide reductase complex), nosZ (nitrous-oxide reductase) and nosA (an outer-membrane protein and OprC homologue). The study also included several genes related to anaerobic or microaerophilic metabolism: napA (encoding the catalytic subunit of the periplasmic nitrate reductase), ccoN (catalytic subunit of the cytochrome-cbb 3 oxidase), hemN (oxygen-independent coproporphyrinogen-III oxidase), an fnr-like regulatory gene, and azu and fdxA (electron carriers azurin and ferredoxin, respectively). Genes necessary for denitrification are concentrated at 20 to 36 min on the P. aeruginosa chromosome, where they form three separate loci, the nir-nor, nar and nos gene clusters. Genomic DNA of Pseudomonas stutzeri ZoBell was also subjected to SpeI restriction and Southern analysis to assign denitrification genes to individual fragments. A homologue of nosA encoding a putative component of the Cu-processing apparatus for nitrous-oxide reductase was identified. In both P. aeruginosa and P. stutzeri there is evidence for the linkage of anr (fnrA) with hemN and ccoN; and for the presence of a napA gene.

Từ khóa


Tài liệu tham khảo

Arai, 1995, Expression of the nir and nor genes for denitrification in Pseudomonas aeruginosa requires a novel CRP/FNR-related transcriptional regulator, DNR, in addition to ANR., FEBS Lett, 371, 73, 10.1016/0014-5793(95)00885-D

de Boer, 1994, Isolation, sequencing and mutational analysis of a gene cluster involved in nitrite reduction in Paracoccus denitrificans, Antonie Leeuwenhoek, 66, 111, 10.1007/BF00871635

de Boer, 1996, Mutational analysis of the nor gene cluster which encodes nitric-oxide reductase from Paracoccus denitrificans, Eur J Biochem, 242, 592, 10.1111/j.1432-1033.1996.0592r.x

Braun, 1991, Marker exchange of the structural genes for nitric oxide reductase blocks the denitrification pathway of Pseudomonas stutzeri at nitric oxide., J Biol Chem, 266, 22785, 10.1016/S0021-9258(18)54420-3

Braun, 1992, The structural genes of the nitric oxide reductase complex from Pseudomonas stutzeri are part of a 30-kilobase gene cluster for denitrification., J Bacteriol, 174, 2394, 10.1128/JB.174.7.2394-2397.1992

Chen, 1993, A simple and rapid method for the preparation of gram-negative bacterial genomic DNA., Nucleic Acids Res, 21, 2260, 10.1093/nar/21.9.2260

Chomczynski, 1992, One-hour downward alkaline capillary transfer for blotting of DNA and RNA., Anal Biochem, 201, 134, 10.1016/0003-2697(92)90185-A

Clark, 1989, A NosA-specific bacteriophage can be used to select denitrification-defective mutants of Pseudomonas stutzeri, J Gen Microbiol, 135, 2569

Cuypers, 1993, Anaerobic control of denitrification in Pseudomonas stutzeri escapes mutagenesis of an fnr-like gene., J Bacteriol, 175, 7236, 10.1128/JB.175.22.7236-7246.1993

Engler-Blum, 1993, Reduction of background problems in non-radioactive Northern and Southern blot analyses enables higher sensitivity than 32P- based hybridizations., Anal Biochem, 210, 235, 10.1006/abio.1993.1189

Farinha, 1993, Localization of the virulence-associated genes pilA, pilR, rpoN, fliA, fliC, ent, and fbp on the physical map of Pseudomonas aeruginosa PAO1 by pulsed-field electrophoresis., Infect Immun, 61, 1571, 10.1128/IAI.61.4.1571-1575.1993

Galimand, 1991, Positive FNR-like control of anaerobic arginine degradation and nitrate respiration in Pseudomonas aeruginosa, J Bacteriol, 173, 1598, 10.1128/JB.173.5.1598-1606.1991

de Gier, 1996, Structural and functional analysis of aa 3-type and cbb 3-type cytochrome c oxidases of Paracoccus denitrificans reveals significant differences in proton-pump design., Mol Microbiol, 20, 1247, 10.1111/j.1365-2958.1996.tb02644.x

Ginard, 1997, Genome organization of Pseudomonas stutzeri and resulting taxonomic and evolutionary considerations., Int J Syst Bacteriol, 47, 132, 10.1099/00207713-47-1-132

Glockner, 1996, Sequence analysis of an internal 9.72-kb segment from the 30-kb denitrification gene cluster of Pseudomonas stutzeri, Biochim Biophys Acta, 1277, 6, 10.1016/S0005-2728(96)00108-9

van Hartingsveldt, 1973, Mapping and characterization of mutants of Pseudomonas aeruginosa affected in nitrate respiration in aerobic or anaerobic growth., J Gen Microbiol, 74, 97, 10.1099/00221287-74-1-97

Hoitink, 1990, Isolation and sequencing of the Alcaligenes denitrificans azurin-encoding gene: comparison with the genes encoding blue copper proteins from Pseudomonas aeruginosa and Alcaligenes faecalis, Gene, 90, 15, 10.1016/0378-1119(90)90434-S

Holloway, 1994, Genomic mapping of Pseudomonas aeruginosa PAO., Microbiology, 140, 2907, 10.1099/13500872-140-11-2907

Jeter, 1984, Chromosomal location and function of genes affecting Pseudomonas aeruginosa nitrate assimilation., J Bacteriol, 157, 673, 10.1128/JB.157.2.673-677.1984

Jüngst, 1991a, Close linkage in Pseudomonas stutzeri of the structural genes for respiratory nitrite reductase and nitrous oxide reductase, and other essential genes for denitrification., Mol Gen Genet, 225, 241, 10.1007/BF00269855

Jüngst, 1991b, The nirSTBM region coding for cytochrome cd 1-dependent nitrite respiration of Pseudomonas stutzeri consists of a cluster of mono-, di-, and tetraheme proteins., FEBS Lett, 279, 205, 10.1016/0014-5793(91)80150-2

Kawasaki, 1997, Gene cluster of dissimilatory nitrite reductase (nir) from Pseudomonas aeruginosa: sequencing and identification of a locus for heme d 1 biosynthesis., J Bacteriol, 179, 235, 10.1128/JB.179.1.235-242.1997

Köhler, 1989, Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions., J Bacteriol, 171, 4326, 10.1128/JB.171.8.4326-4333.1989

Lee, 1991, Molecular characterization of nosA, a Pseudomonas stutzeri gene encoding an outer membrane protein required to make copper-containing N2O reductase., J Bacteriol, 173, 5406, 10.1128/JB.173.17.5406-5413.1991

Liao, 1996, Physical mapping of 32 genetic markers on the Pseudomonas aeruginosa PAO1 chromosome., Microbiology, 142, 79, 10.1099/13500872-142-1-79

Meade, 1982, Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis., J Bacteriol, 149, 114, 10.1128/JB.149.1.114-122.1982

Mokhele, 1987, A Pseudomonas stutzeri outer membrane protein inserts copper into N2O reductase., J Bacteriol, 169, 5721, 10.1128/JB.169.12.5721-5726.1987

Nordling, 1990, The structural gene for cytochrome c 551 from Pseudomonas aeruginosa: the nucleotide sequence shows a location down-stream of the nitrite reductase gene., FEBS Lett, 259, 230, 10.1016/0014-5793(90)80015-B

Nunoshiba, 1993, Activation by nitric oxide of an oxidative-stress response that defends Escherichia coli against activated macrophages., Proc Natl Acad Sci USA, 90, 9993, 10.1073/pnas.90.21.9993

Preisig, 1993, Genes for a microaerobically induced oxidase complex in Bradyrhizobium japonicum are essential for a nitrogen-fixing endosymbiosis., Proc Natl Acad Sci USA, 90, 3309, 10.1073/pnas.90.8.3309

Römling, 1991, The impact of two-dimensional pulsed-field gel electrophoresis techniques for the consistent and complete mapping of bacterial genomes: refined physical map of Pseudomonas aeruginosa PAO., Nucleic Acids Res, 19, 3199, 10.1093/nar/19.12.3199

Römling, 1994, Bacterial genome analysis by pulsed field gel electrophoresis techniques., Adv Electrophor, 7, 355

Saeki, 1988, Pseudomonas stutzeri ferredoxin: close similarity to Azotobacter vinelandii and Pseudomonas ovalis ferredoxins., J Biochem, 104, 242, 10.1093/oxfordjournals.jbchem.a122450

Sambrook, 1989, Molecular Cloning: a Laboratory Manual

Sawers, 1991, Identification and molecular characterization of a transcriptional regulator from Pseudomonas aeruginosa PAO1 exhibiting structural and functional similarity to the FNR protein of Escherichia coli, Mol Microbiol, 5, 1469, 10.1111/j.1365-2958.1991.tb00793.x

Sias, 1980, The assimilatory and dissimilatory nitrate reductases of Pseudomonas aeruginosa are encoded by different genes., J Gen Microbiol, 118, 229

Siddiqui, 1993, Structure and function of a periplasmic nitrate reductase in Alcaligenes eutrophus H16., J Bacteriol, 175, 5867, 10.1128/JB.175.18.5867-5876.1993

Sodergren, 1988, narI region of the Escherichia coli nitrate reductase (nar) operon contains two genes., J Bacteriol, 170, 1721, 10.1128/JB.170.4.1721-1729.1988

Viebrock, 1988, Molecular cloning, heterologous expression, and primary structure of the structural gene for the copper enzyme nitrous oxide reductase from denitrifying Pseudomonas stutzeri, J Bacteriol, 170, 4658, 10.1128/JB.170.10.4658-4668.1988

Vijgenboom, 1995, Physiological role and expression of the blue copper protein azurin in Pseudomonas aeruginosa, J Inorg Biochem, 59, 720, 10.1016/0162-0134(95)97808-4

Vollack, 1996, Mapping of denitrification genes from Pseudomonas aeruginosa by macro-restriction analysis., Biospectrum, 1996

Yannone, 1997, Identification of a palindromic sequence that is responsible for the up-regulation of NADPH-ferredoxin reductase in a ferredoxin I deletion strain of Azotobacter vinelandii, J Biol Chem, 272, 14454, 10.1074/jbc.272.22.14454

Ye, 1995, Anaerobic activation of the entire denitrification pathway in Pseudomonas aeruginosa requires Anr, an analog of Fnr., J Bacteriol, 177, 3606, 10.1128/JB.177.12.3606-3609.1995

Yoneyama, 1996, Protein C (OprC) of the outer membrane of Pseudomonas aeruginosa is a copper-regulated channel protein., Microbiology, 142, 2137, 10.1099/13500872-142-8-2137

Zumft, 1997, Cell biology and the molecular basis of denitrification., Microbiol Mol Biol Rev, 61, 533, 10.1128/.61.4.533-616.1997

Zumft, 1992, Derived amino acid sequences of the nosZ gene (respiratory N2O reductase) from Alcaligenes eutrophus, Pseudomonas aeruginosa and Pseudomonas stutzeri reveal potential copper-binding residues: implications for the CuA site of N2O reductase and cytochrome-c oxidase., Eur J Biochem, 208, 31, 10.1111/j.1432-1033.1992.tb17156.x

Zumft, 1994, Nitric oxide reductase from Pseudomonas stutzeri: primary structure and gene organization of a novel bacterial cytochrome bc complex., Eur J Biochem, 219, 481, 10.1111/j.1432-1033.1994.tb19962.x