Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Apr;177(8):2143–2150. doi: 10.1128/jb.177.8.2143-2150.1995

Genes encoded on a cyanobacterial plasmid are transcriptionally regulated by sulfur availability and CysR.

M L Nicholson 1, D E Laudenbach 1
PMCID: PMC176859  PMID: 7536734

Abstract

A cyanobacterial sulfur-regulated gene (cysR), which encodes a protein with similarity to the Crp family of prokaryotic regulatory proteins, has recently been isolated and characterized. Polyacrylamide gel electrophoresis of periplasmic protein extracts reveals that a cysR mutant fails to synthesize a 36-kDa polypeptide that is normally induced in wild-type cells that have been grown under sulfur-deficient conditions. The amino-terminal sequence of this protein was obtained, and a synthetic oligonucleotide was used to isolated a clone containing a 1.9-kb NruI-KpnI fragment from a Synechococcus sp. strain PCC 7942 genomic library. RNA blot analysis indicates that this fragment encodes a transcript that is detectable in wild-type but not cysR mutant cells that have been starved for sulfur. DNA blot analysis revealed that the 1.9-kb NruI-KpnI fragment is contained within the Ba4 BamHI fragment of the endogenous 50-kb plasmid pANL. RNA blot studies indicate that the accumulation of a large number of pANL transcripts is regulated by sulfur levels and CysR. DNA sequence analysis confirmed that the gene encoding the sulfur-regulated 36-kDa periplasmic protein is encoded on the Ba4 fragment of pANL. The sequence of the 36-kDa protein displays sequence similarity to the enzyme catalase, and two downstream proteins exhibit 25 and 62% identity to a subunit of a P-type ATPase complex involved in Mg2+ transport and a chromate resistance determinant, respectively. Surprisingly, a strain in which the putative chromate resistance gene was interrupted by a drug resistance marker exhibited increased resistance to chromate when grown in media containing low sulfate concentrations. The possible role of this protein in the acclimation of cyanobacteria to conditions of low sulfur availability is discussed.

Full Text

The Full Text of this article is available as a PDF (366.6 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Batut J., Daveran-Mingot M. L., David M., Jacobs J., Garnerone A. M., Kahn D. fixK, a gene homologous with fnr and crp from Escherichia coli, regulates nitrogen fixation genes both positively and negatively in Rhizobium meliloti. EMBO J. 1989 Apr;8(4):1279–1286. doi: 10.1002/j.1460-2075.1989.tb03502.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beckman D. L., Trawick D. R., Kranz R. G. Bacterial cytochromes c biogenesis. Genes Dev. 1992 Feb;6(2):268–283. doi: 10.1101/gad.6.2.268. [DOI] [PubMed] [Google Scholar]
  4. Cervantes C., Ohtake H., Chu L., Misra T. K., Silver S. Cloning, nucleotide sequence, and expression of the chromate resistance determinant of Pseudomonas aeruginosa plasmid pUM505. J Bacteriol. 1990 Jan;172(1):287–291. doi: 10.1128/jb.172.1.287-291.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ebright R. H., Cossart P., Gicquel-Sanzey B., Beckwith J. Mutations that alter the DNA sequence specificity of the catabolite gene activator protein of E. coli. Nature. 1984 Sep 20;311(5983):232–235. doi: 10.1038/311232a0. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Green L. S., Grossman A. R. Changes in sulfate transport characteristics and protein composition of Anacystis nidulans R2 during sulfur deprivation. J Bacteriol. 1988 Feb;170(2):583–587. doi: 10.1128/jb.170.2.583-587.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Green L. S., Laudenbach D. E., Grossman A. R. A region of a cyanobacterial genome required for sulfate transport. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1949–1953. doi: 10.1073/pnas.86.6.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jeanjean R., Broda E. Dependence of sulphate uptake by Anacystis nidulans on energy, on osmotic shock and on sulphate stravation. Arch Microbiol. 1977 Jul 26;114(1):19–23. doi: 10.1007/BF00429625. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lau R. H., Doolittle W. F. Covalently closed circular DNAs in closely related unicellular cyanobacteria. J Bacteriol. 1979 Jan;137(1):648–652. doi: 10.1128/jb.137.1.648-652.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lau R. H., Sapienza C., Doolittle W. F. Cyanobacterial plasmids: their widespread occurrence, and the existence of regions of homology between plasmids in the same and different species. Mol Gen Genet. 1980 Apr;178(1):203–211. doi: 10.1007/BF00267230. [DOI] [PubMed] [Google Scholar]
  13. Laudenbach D. E., Ehrhardt D., Green L., Grossman A. Isolation and characterization of a sulfur-regulated gene encoding a periplasmically localized protein with sequence similarity to rhodanese. J Bacteriol. 1991 May;173(9):2751–2760. doi: 10.1128/jb.173.9.2751-2760.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laudenbach D. E., Grossman A. R. Characterization and mutagenesis of sulfur-regulated genes in a cyanobacterium: evidence for function in sulfate transport. J Bacteriol. 1991 May;173(9):2739–2750. doi: 10.1128/jb.173.9.2739-2750.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laudenbach D. E., Herbert S. K., McDowell C., Fork D. C., Grossman A. R., Straus N. A. Cytochrome c-553 is not required for photosynthetic activity in the cyanobacterium Synechococcus. Plant Cell. 1990 Sep;2(9):913–924. doi: 10.1105/tpc.2.9.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laudenbach D. E., Reith M. E., Straus N. A. Isolation, sequence analysis, and transcriptional studies of the flavodoxin gene from Anacystis nidulans R2. J Bacteriol. 1988 Jan;170(1):258–265. doi: 10.1128/jb.170.1.258-265.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liesegang H., Lemke K., Siddiqui R. A., Schlegel H. G. Characterization of the inducible nickel and cobalt resistance determinant cnr from pMOL28 of Alcaligenes eutrophus CH34. J Bacteriol. 1993 Feb;175(3):767–778. doi: 10.1128/jb.175.3.767-778.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nies A., Nies D. H., Silver S. Nucleotide sequence and expression of a plasmid-encoded chromate resistance determinant from Alcaligenes eutrophus. J Biol Chem. 1990 Apr 5;265(10):5648–5653. [PubMed] [Google Scholar]
  19. Pardee A. B., Prestidge L. S., Whipple M. B., Dreyfuss J. A binding site for sulfate and its relation to sulfate transport into Salmonella typhimurium. J Biol Chem. 1966 Sep 10;241(17):3962–3969. [PubMed] [Google Scholar]
  20. Pardee A. B. Purification and properties of a sulfate-binding protein from Salmonella typhimurium. J Biol Chem. 1966 Dec 25;241(24):5886–5892. [PubMed] [Google Scholar]
  21. Pastan I., Adhya S. Cyclic adenosine 5'-monophosphate in Escherichia coli. Bacteriol Rev. 1976 Sep;40(3):527–551. doi: 10.1128/br.40.3.527-551.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  23. Reith M. E., Laudenbach D. E., Straus N. A. Isolation and nucleotide sequence analysis of the ferredoxin I gene from the cyanobacterium Anacystis nidulans R2. J Bacteriol. 1986 Dec;168(3):1319–1324. doi: 10.1128/jb.168.3.1319-1324.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Snavely M. D., Miller C. G., Maguire M. E. The mgtB Mg2+ transport locus of Salmonella typhimurium encodes a P-type ATPase. J Biol Chem. 1991 Jan 15;266(2):815–823. [PubMed] [Google Scholar]
  26. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tandeau de Marsac N., Borrias W. E., Kuhlemeier C. J., Castets A. M., van Arkel G. A., van den Hondel C. A. A new approach for molecular cloning in cyanobacteria: cloning of an Anacystis nidulans met gene using a Tn901-induced mutant. Gene. 1982 Nov;20(1):111–119. doi: 10.1016/0378-1119(82)90092-0. [DOI] [PubMed] [Google Scholar]
  28. Tomioka N., Sugiura M. The complete nucleotide sequence of a 16S ribosomal RNA gene from a blue-green alga, Anacystis nidulans. Mol Gen Genet. 1983;191(1):46–50. doi: 10.1007/BF00330888. [DOI] [PubMed] [Google Scholar]
  29. Unden G., Guest J. R. Isolation and characterization of the Fnr protein, the transcriptional regulator of anaerobic electron transport in Escherichia coli. Eur J Biochem. 1985 Jan 2;146(1):193–199. doi: 10.1111/j.1432-1033.1985.tb08638.x. [DOI] [PubMed] [Google Scholar]
  30. Vega-Palas M. A., Flores E., Herrero A. NtcA, a global nitrogen regulator from the cyanobacterium Synechococcus that belongs to the Crp family of bacterial regulators. Mol Microbiol. 1992 Jul;6(13):1853–1859. doi: 10.1111/j.1365-2958.1992.tb01357.x. [DOI] [PubMed] [Google Scholar]
  31. Wei T. F., Ramasubramanian T. S., Pu F., Golden J. W. Anabaena sp. strain PCC 7120 bifA gene encoding a sequence-specific DNA-binding protein cloned by in vivo transcriptional interference selection. J Bacteriol. 1993 Jul;175(13):4025–4035. doi: 10.1128/jb.175.13.4025-4035.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. van den Hondel C. A., Keegstra W., Borrias W. E., van Arkel G. A. Homology of plasmids in strains of unicellular Cyanobacteria. Plasmid. 1979 Jul;2(3):323–333. doi: 10.1016/0147-619x(79)90016-7. [DOI] [PubMed] [Google Scholar]
  33. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. von Ossowski I., Hausner G., Loewen P. C. Molecular evolutionary analysis based on the amino acid sequence of catalase. J Mol Evol. 1993 Jul;37(1):71–76. doi: 10.1007/BF00170464. [DOI] [PubMed] [Google Scholar]

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

RESOURCES