Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Dec;179(23):7343–7350. doi: 10.1128/jb.179.23.7343-7350.1997

A cysG mutant strain of Rhizobium etli pleiotropically defective in sulfate and nitrate assimilation.

R Tate 1, A Riccio 1, M Iaccarino 1, E J Patriarca 1
PMCID: PMC179684  PMID: 9393698

Abstract

By its inability to grow on sulfate as the sole sulfur source, a mutant strain (CTNUX8) of Rhizobium etli carrying Tn5 was isolated and characterized. Sequence analysis showed that Tn5 is inserted into a cysG (siroheme synthetase)-homologous gene. By RNase protection assays, it was established that the cysG-like gene had a basal level of expression in thiosulfate- or cysteine-grown cells, which was induced when sulfate or methionine was used. Unlike its wild-type parent (strain CE3), the mutant strain, CTNUX8, was also unable to grow on nitrate as the sole nitrogen source and was unable to induce a high level of nitrite reductase. Despite its pleiotropic phenotype, strain CTNUX8 was able to induce pink, effective (N2-fixing) nodules on the roots of Phaseolus vulgaris plants. However, mixed inoculation experiments showed that strain CTNUX8 is significantly different from the wild type in its ability to nodulate. Our data support the notion that sulfate (or sulfite) is the sulfur source of R. etli in the rhizosphere, while cysteine, methionine, or glutathione is supplied by the root cells to bacteria growing inside the plant.

Full Text

The Full Text of this article is available as a PDF (375.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blanche F., Robin C., Couder M., Faucher D., Cauchois L., Cameron B., Crouzet J. Purification, characterization, and molecular cloning of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Methanobacterium ivanovii. J Bacteriol. 1991 Aug;173(15):4637–4645. doi: 10.1128/jb.173.15.4637-4645.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cole J. A., Newman B. M., White P. Biochemical and genetic characterization of nirB mutants of Escherichia coli K 12 pleiotropically defective in nitrite and sulphite reduction. J Gen Microbiol. 1980 Oct;120(2):475–483. doi: 10.1099/00221287-120-2-475. [DOI] [PubMed] [Google Scholar]
  3. DEVITO P. C., DREYFUSS J. METABOLIC REGULATION OF ADENOSINE TRIPHOSPHATE SULFURYLASE IN YEAST. J Bacteriol. 1964 Nov;88:1341–1348. doi: 10.1128/jb.88.5.1341-1348.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fazzio T. G., Roth J. R. Evidence that the CysG protein catalyzes the first reaction specific to B12 synthesis in Salmonella typhimurium, insertion of cobalt. J Bacteriol. 1996 Dec;178(23):6952–6959. doi: 10.1128/jb.178.23.6952-6959.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gage D. J., Bobo T., Long S. R. Use of green fluorescent protein to visualize the early events of symbiosis between Rhizobium meliloti and alfalfa (Medicago sativa). J Bacteriol. 1996 Dec;178(24):7159–7166. doi: 10.1128/jb.178.24.7159-7166.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goldman B. S., Roth J. R. Genetic structure and regulation of the cysG gene in Salmonella typhimurium. J Bacteriol. 1993 Mar;175(5):1457–1466. doi: 10.1128/jb.175.5.1457-1466.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hansen J., Muldbjerg M., Chérest H., Surdin-Kerjan Y. Siroheme biosynthesis in Saccharomyces cerevisiae requires the products of both the MET1 and MET8 genes. FEBS Lett. 1997 Jan 13;401(1):20–24. doi: 10.1016/s0014-5793(96)01423-8. [DOI] [PubMed] [Google Scholar]
  8. Lerouge P., Roche P., Faucher C., Maillet F., Truchet G., Promé J. C., Dénarié J. Symbiotic host-specificity of Rhizobium meliloti is determined by a sulphated and acylated glucosamine oligosaccharide signal. Nature. 1990 Apr 19;344(6268):781–784. doi: 10.1038/344781a0. [DOI] [PubMed] [Google Scholar]
  9. Murphy M. J., Siegel L. M., Tove S. R., Kamin H. Siroheme: a new prosthetic group participating in six-electron reduction reactions catalyzed by both sulfite and nitrite reductases. Proc Natl Acad Sci U S A. 1974 Mar;71(3):612–616. doi: 10.1073/pnas.71.3.612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nakamura T., Iwahashi H., Eguchi Y. Enzymatic proof for the identity of the S-sulfocysteine synthase and cysteine synthase B of Salmonella typhimurium. J Bacteriol. 1984 Jun;158(3):1122–1127. doi: 10.1128/jb.158.3.1122-1127.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Patriarca E. J., Riccio A., Taté R., Colonna-Romano S., Iaccarino M., Defez R. The ntrBC genes of Rhizobium leguminosarum are part of a complex operon subject to negative regulation. Mol Microbiol. 1993 Aug;9(3):569–577. doi: 10.1111/j.1365-2958.1993.tb01717.x. [DOI] [PubMed] [Google Scholar]
  12. Roth J. R., Lawrence J. G., Rubenfield M., Kieffer-Higgins S., Church G. M. Characterization of the cobalamin (vitamin B12) biosynthetic genes of Salmonella typhimurium. J Bacteriol. 1993 Jun;175(11):3303–3316. doi: 10.1128/jb.175.11.3303-3316.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Schwedock J. S., Long S. R. Rhizobium meliloti genes involved in sulfate activation: the two copies of nodPQ and a new locus, saa. Genetics. 1992 Dec;132(4):899–909. doi: 10.1093/genetics/132.4.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schwedock J., Long S. R. ATP sulphurylase activity of the nodP and nodQ gene products of Rhizobium meliloti. Nature. 1990 Dec 13;348(6302):644–647. doi: 10.1038/348644a0. [DOI] [PubMed] [Google Scholar]
  15. Segovia L., Young J. P., Martínez-Romero E. Reclassification of American Rhizobium leguminosarum biovar phaseoli type I strains as Rhizobium etli sp. nov. Int J Syst Bacteriol. 1993 Apr;43(2):374–377. doi: 10.1099/00207713-43-2-374. [DOI] [PubMed] [Google Scholar]
  16. Siegel L. M., Murphy M. J., Kamin H. Reduced nicotinamide adenine dinucleotide phosphate-sulfite reductase of enterobacteria. I. The Escherichia coli hemoflavoprotein: molecular parameters and prosthetic groups. J Biol Chem. 1973 Jan 10;248(1):251–264. [PubMed] [Google Scholar]
  17. Spencer J. B., Stolowich N. J., Roessner C. A., Scott A. I. The Escherichia coli cysG gene encodes the multifunctional protein, siroheme synthase. FEBS Lett. 1993 Nov 29;335(1):57–60. doi: 10.1016/0014-5793(93)80438-z. [DOI] [PubMed] [Google Scholar]
  18. Thomas D., Cherest H., Surdin-Kerjan Y. Elements involved in S-adenosylmethionine-mediated regulation of the Saccharomyces cerevisiae MET25 gene. Mol Cell Biol. 1989 Aug;9(8):3292–3298. doi: 10.1128/mcb.9.8.3292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Triplett E. W., Sadowsky M. J. Genetics of competition for nodulation of legumes. Annu Rev Microbiol. 1992;46:399–428. doi: 10.1146/annurev.mi.46.100192.002151. [DOI] [PubMed] [Google Scholar]
  20. Vega J. M., Guerrero M. G., Leadbetter E., Losada M. Reduced nicotinamide-adenine dinucleotide-nitrite reductase from Azotobacter chroococcum. Biochem J. 1973 Aug;133(4):701–708. doi: 10.1042/bj1330701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Verma DPS. Signals in Root Nodule Organogenesis and Endocytosis of Rhizobium. Plant Cell. 1992 Apr;4(4):373–382. doi: 10.1105/tpc.4.4.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weinstein M., Roberts R. C., Helinski D. R. A region of the broad-host-range plasmid RK2 causes stable in planta inheritance of plasmids in Rhizobium meliloti cells isolated from alfalfa root nodules. J Bacteriol. 1992 Nov;174(22):7486–7489. doi: 10.1128/jb.174.22.7486-7489.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Woodcock D. M., Crowther P. J., Doherty J., Jefferson S., DeCruz E., Noyer-Weidner M., Smith S. S., Michael M. Z., Graham M. W. Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants. Nucleic Acids Res. 1989 May 11;17(9):3469–3478. doi: 10.1093/nar/17.9.3469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wu J. Y., Siegel L. M., Kredich N. M. High-level expression of Escherichia coli NADPH-sulfite reductase: requirement for a cloned cysG plasmid to overcome limiting siroheme cofactor. J Bacteriol. 1991 Jan;173(1):325–333. doi: 10.1128/jb.173.1.325-333.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. van Rhijn P., Vanderleyden J. The Rhizobium-plant symbiosis. Microbiol Rev. 1995 Mar;59(1):124–142. doi: 10.1128/mr.59.1.124-142.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES