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. 1989 Feb;171(2):1093–1099. doi: 10.1128/jb.171.2.1093-1099.1989

An rpoN-like gene of Alcaligenes eutrophus and Pseudomonas facilis controls expression of diverse metabolic pathways, including hydrogen oxidation.

D Römermann 1, J Warrelmann 1, R A Bender 1, B Friedrich 1
PMCID: PMC209706  PMID: 2536672

Abstract

Pleiotropic mutants of Alcaligenes eutrophus with the phenotype Hno- have been characterized previously. They are deficient in several diverse metabolic activities, including hydrogen oxidation, nitrate and urea assimilation, denitrification, and various substrate transport systems. Phenotypically similar mutants were identified among hydrogenase-deficient strains of Pseudomonas facilis. The Tn5-labeled hno gene was cloned from a genomic DNA library of A. eutrophus and used to identify the corresponding unimpaired wild-type DNA sequence. The recombinant plasmid pCH148 contained an insert of 12.3 kilobase pairs and was shown to restore the Hno+ phenotype to mutants of A. eutrophus and P. facilis. A cosmid isolated from a DNA library of P. facilis also exhibited intergeneric Hno-complementing activity. The cloned hno loci from both organisms showed DNA homology by Southern blot hybridization. A subclone of pCH148 which contained a 6.5-kilobase-pair insert was constructed. The resulting hybrid, pCH170, not only was able to complement Hno- mutants but also relieved glutamine auxotrophy in NtrA- mutants of enteric bacteria. This suggests that the hno gene product from A. eutrophus is functionally similar to the NtrA protein, which has been identified as a novel sigma factor (sigma 54) of RNA polymerase.

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Selected References

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  1. Alvarez-Morales A., Hennecke H. Expression of Rhizobium japonicum nifH and nifDK operons can be activated by the Klebsiella pneumonia nifA protein but not by the product of ntrC. Mol Gen Genet. 1985;199(2):306–314. doi: 10.1007/BF00330273. [DOI] [PubMed] [Google Scholar]
  2. Bender R. A., Eades L. J. A nonsense mutation in the structural gene for glutamine synthetase leading to loss of nitrogen regulation in Klebsiella aerogenes. Mol Gen Genet. 1982;187(3):414–418. doi: 10.1007/BF00332621. [DOI] [PubMed] [Google Scholar]
  3. Birkmann A., Sawers R. G., Böck A. Involvement of the ntrA gene product in the anaerobic metabolism of Escherichia coli. Mol Gen Genet. 1987 Dec;210(3):535–542. doi: 10.1007/BF00327209. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bowien B., Schlegel H. G. Physiology and biochemistry of aerobic hydrogen-oxidizing bacteria. Annu Rev Microbiol. 1981;35:405–452. doi: 10.1146/annurev.mi.35.100181.002201. [DOI] [PubMed] [Google Scholar]
  6. Eberz G., Hogrefe C., Kortlüke C., Kamienski A., Friedrich B. Molecular cloning of structural and regulatory hydrogenase (hox) genes of Alcaligenes eutrophus H16. J Bacteriol. 1986 Nov;168(2):636–641. doi: 10.1128/jb.168.2.636-641.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Errington J. Microbiology. Regulation of sporulation. Nature. 1988 Jun 2;333(6172):399–400. doi: 10.1038/333399a0. [DOI] [PubMed] [Google Scholar]
  8. Fischer H M, Hennecke H. Direct response of Bradyrhizobium japonicum nifA-mediated nif gene regulation to cellular oxygen status. Mol Gen Genet. 1987 Oct;209(3):621–626. doi: 10.1007/BF00331174. [DOI] [PubMed] [Google Scholar]
  9. Friedrich B., Heine E., Finck A., Friedrich C. G. Nickel requirement for active hydrogenase formation in Alcaligenes eutrophus. J Bacteriol. 1981 Mar;145(3):1144–1149. doi: 10.1128/jb.145.3.1144-1149.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friedrich B., Hogrefe C., Schlegel H. G. Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol. 1981 Jul;147(1):198–205. doi: 10.1128/jb.147.1.198-205.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Friedrich C. G., Friedrich B. Regulation of hydrogenase formation is temperature sensitive and plasmid coded in Alcaligenes eutrophus. J Bacteriol. 1983 Jan;153(1):176–181. doi: 10.1128/jb.153.1.176-181.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grossman A. D., Erickson J. W., Gross C. A. The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell. 1984 Sep;38(2):383–390. doi: 10.1016/0092-8674(84)90493-8. [DOI] [PubMed] [Google Scholar]
  13. Hirschman J., Wong P. K., Sei K., Keener J., Kustu S. Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7525–7529. doi: 10.1073/pnas.82.22.7525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hogrefe C., Friedrich B. Isolation and characterization of megaplasmid DNA from lithoautotrophic bacteria. Plasmid. 1984 Nov;12(3):161–169. doi: 10.1016/0147-619x(84)90040-4. [DOI] [PubMed] [Google Scholar]
  15. Hogrefe C., Römermann D., Friedrich B. Alcaligenes eutrophus hydrogenase genes (Hox). J Bacteriol. 1984 Apr;158(1):43–48. doi: 10.1128/jb.158.1.43-48.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hunt T. P., Magasanik B. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453–8457. doi: 10.1073/pnas.82.24.8453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inouye S., Ebina Y., Nakazawa A., Nakazawa T. Nucleotide sequence surrounding transcription initiation site of xylABC operon on TOL plasmid of Pseudomonas putida. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1688–1691. doi: 10.1073/pnas.81.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnson K., Parker M. L., Lory S. Nucleotide sequence and transcriptional initiation site of two Pseudomonas aeruginosa pilin genes. J Biol Chem. 1986 Nov 25;261(33):15703–15708. [PubMed] [Google Scholar]
  19. Klintworth R., Husemann M., Salnikow J., Bowien B. Chromosomal and plasmid locations for phosphoribulokinase genes in Alcaligenes eutrophus. J Bacteriol. 1985 Nov;164(2):954–956. doi: 10.1128/jb.164.2.954-956.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Lambden P. R., Guest J. R. Mutants of Escherichia coli K12 unable to use fumarate as an anaerobic electron acceptor. J Gen Microbiol. 1976 Dec;97(2):145–160. doi: 10.1099/00221287-97-2-145. [DOI] [PubMed] [Google Scholar]
  23. MacNeil T., MacNeil D., Tyler B. Fine-structure deletion map and complementation analysis of the glnA-glnL-glnG region in Escherichia coli. J Bacteriol. 1982 Jun;150(3):1302–1313. doi: 10.1128/jb.150.3.1302-1313.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  25. Merrick M. J., Stewart W. D. Studies on the regulation and function of the Klebsiella pneumoniae ntrA gene. Gene. 1985;35(3):297–303. doi: 10.1016/0378-1119(85)90008-3. [DOI] [PubMed] [Google Scholar]
  26. Merrick M., Gibbins J., Toukdarian A. The nucleotide sequence of the sigma factor gene ntrA (rpoN) of Azotobacter vinelandii: analysis of conserved sequences in NtrA proteins. Mol Gen Genet. 1987 Dec;210(2):323–330. doi: 10.1007/BF00325701. [DOI] [PubMed] [Google Scholar]
  27. O'Neill E. A., Kiely G. M., Bender R. A. Transposon Tn5 encodes streptomycin resistance in nonenteric bacteria. J Bacteriol. 1984 Jul;159(1):388–389. doi: 10.1128/jb.159.1.388-389.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reitzer L. J., Bueno R., Cheng W. D., Abrams S. A., Rothstein D. M., Hunt T. P., Tyler B., Magasanik B. Mutations that create new promoters suppress the sigma 54 dependence of glnA transcription in Escherichia coli. J Bacteriol. 1987 Sep;169(9):4279–4284. doi: 10.1128/jb.169.9.4279-4284.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ronson C. W., Nixon B. T., Albright L. M., Ausubel F. M. Rhizobium meliloti ntrA (rpoN) gene is required for diverse metabolic functions. J Bacteriol. 1987 Jun;169(6):2424–2431. doi: 10.1128/jb.169.6.2424-2431.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Römermann D., Friedrich B. Denitrification by Alcaligenes eutrophus is plasmid dependent. J Bacteriol. 1985 May;162(2):852–854. doi: 10.1128/jb.162.2.852-854.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. SCHLEGEL H. G., KALTWASSER H., GOTTSCHALK G. [A submersion method for culture of hydrogen-oxidizing bacteria: growth physiological studies]. Arch Mikrobiol. 1961;38:209–222. [PubMed] [Google Scholar]
  32. Simon R. High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet. 1984;196(3):413–420. doi: 10.1007/BF00436188. [DOI] [PubMed] [Google Scholar]
  33. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  34. Streicher S. L., Bender R. A., Magasanik B. Genetic control of glutamine synthetase in Klebiella aerogenes. J Bacteriol. 1975 Jan;121(1):320–331. doi: 10.1128/jb.121.1.320-331.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thöny B., Fischer H. M., Anthamatten D., Bruderer T., Hennecke H. The symbiotic nitrogen fixation regulatory operon (fixRnifA) of Bradyrhizobium japonicum is expressed aerobically and is subject to a novel, nifA-independent type of activation. Nucleic Acids Res. 1987 Oct 26;15(20):8479–8499. doi: 10.1093/nar/15.20.8479. [DOI] [PMC free article] [PubMed] [Google Scholar]

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