Skip to main content
SpringerLink
Log in

Osmotic and growth-phase dependent regulation of the eta gene of Staphylococcus aureus: a role for DNA supercoiling

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Transcriptional fusions were constructed between the promoter for the epidermolytic toxin A (eta) gene of Staphylococcus aureus and the luxAB and xylE reporter gene systems. The expression of the fusion products was found to be dependent upon the accessory gene regulator (agr) locus and was observed to increase significantly during the transition from the exponential to the stationary phase of growth. Furthermore the expression of the eta gene promoter was found to be osmotically regulated, with the expression levels of the eta fusions being inversely related to the osmolyte levels. The ability of environmental factors to influence DNA topology (and thence gene expression) was investigated. High osmolarity (0.7 M NaCl) resulted in an increase in the degree of negative supercoiling of plasmid DNA in the S. aureus strain 8325-4 (Agr+) but not in strain ISP546 (Agr−). Furthermore the eta promoter was strongly induced in S. aureus cultures grown in the presence of sub-inhibitory concentrations of novobiocin, a DNA gyrase inhibitor. However this induction was independent of agr, suggesting that the eta promoter is subject to both agr-dependent (osmolarity, growth phase) and -independent (DNA topology) regulatory processes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ahmed KA, Stewart GSAB (1988) Cloning of the lux genes into Lactobacillus casei and Streptococcus lactis: phosphate-dependent light production. Biochem Soc Trans 16:1068

    Google Scholar 

  • Ahmed KA, Stewart GSAB (1991) The production of bioluminescent lactic acid bacteria suitable for the rapid assessment of starter culture activity in milk. J Appl Bacteriol 70:113–120

    Google Scholar 

  • Balke V, Gralla J (1987) Changes in the linking number of super-coiled DNA accompany growth transitions in E. coli. J Bacteriol 169:4499–4506

    Google Scholar 

  • Bayles KW, Iandolo JJ (1989) Genetic and molecular analysis of the gene encoding staphylococcal enterotoxin D. J Bacteriol 171:4799–4806

    Google Scholar 

  • Bernardini ML, Fontaine A, Sansonetti PJ (1990) The two component regulatory system OmpR-EnvZ controls the virulence of Shigella flexneri. J Bacteriol 172:6274–6281

    Google Scholar 

  • Bjorklind A, Arvidson S (1980) Mutants of Staphylococcus aureus affected in the regulation of exoprotein synthesis. FEMS Lett 7:203–206

    Google Scholar 

  • Chatfield SN, Dorman CJ, Hayward C, Dougan G (1991) Role of ompR dependent genes in Salmonella typhimurium virulence: mutants deficient in OmpC and OmpF are attenuated in vivo. Infect Immun 59:449–452

    Google Scholar 

  • Dorman CJ (1991) DNA supercoiling and environment regulation of gene expression in pathogenic bacteria. Infect Immun 59:745–749

    Google Scholar 

  • Dorman CJ, Barr GC, Ni Bhriain N, Higgins CF (1988) DNA supercoiling and the anaerobic and growth phase regulation of tonB expression. J Bacteriol 170:2816–2826

    Google Scholar 

  • Dorman CJ, Chatfield S, Higgins CF, Hayward C, Dougan G (1989) Characterisation of porin and ompR mutants of a virulent strain of Salmonella typhimurium: ompR mutants are attenuated in vivo. Infect Immun 57:2136–2140

    Google Scholar 

  • Drlica K (1990) Bacterial topoisomerases and the control of DNA supercoiling. Trends Genet 6:433–437

    Google Scholar 

  • Easmon CSF, Adlam C (eds) (1983) Staphylococci and staphylococcal infections. vols 1 and 2. Academic Press, London

    Google Scholar 

  • Galan JE, Curtiss R III (1990) Expression of Salmonella typhimurium genes required for invasion is regulated by changes in DNA supercoiling. Infect Immun 58:1879–1885

    Google Scholar 

  • Gaskill ME, Khan SA (1988) Regulation of enterotoxin B gene in Staphylococcus aureus. J Biol Chem 263:6276–6280

    Google Scholar 

  • Gasson MJ, Anderson PH (1985) High copy number plasmid vectors for use in lactic streptococci. FEMS Lett 30:193–196

    Google Scholar 

  • Higgins CF, Dorman CJ, Stirling DA, Waddell L, Booth IR, May G, Bremer E (1988) A physiological role for DNA supercoiling in the osmotic regulation of gene expression in Salmonella typhimurium and E. coli. Cell 52:569–584

    Google Scholar 

  • Hopewell R, Oram M, Briesewitz R, Fischer LM (1990) DNA cloning and organisation of the Staphylococcus aureus gyrA and gyrB genes: close homology among proteins and implications for 4-quinolone action and resistance. J Bacteriol 172:3481–3484

    Google Scholar 

  • Janzon L, Arvidson S (1990) The role of the δ-lysin gene (hld) in the regulation of virulence genes by the accessory gene regulator (agr) of Staphylococcus aureus. EMBO J 9:1391–1399

    Google Scholar 

  • Janzon L, Lofdahl S, Arvidson S (1986) Evidence for a coordinate transcriptional control of alpha-toxin and protein A synthesis in Staphylococcus aureus. FEMS Lett 33:193–198

    Google Scholar 

  • Janzon L, Lofdahl S, Arvidson S (1989) Identification and nucleotide sequence of the delta-lysin gene, hld, adjacent to the accessory gene regulator (agr) of Staphylococcus aureus. Mol Gen Genet 219:480–485

    Google Scholar 

  • Kreiswirth B, Lofdahl S, Betley MJ, O'Reilly M, Schlievert PM, Bergdoll MS, Novick RP (1983) The toxic shock syndrome exotoxin structural gene is not transmitted by a prophage. Nature 305:709–712

    Google Scholar 

  • Lee CY, Schmidt JJ, Johnson-Winegar AD, Spero L, Iandolo JJ (1987) Sequence determination and comparison of the exfoliative toxin A and toxin B genes from Staphylococcus aureus. J Bacteriol 169:3904–3909

    Google Scholar 

  • Mahmood R, Khan SA (1990) Role of upstream sequences in the expression of the staphylococcal enterotoxin B gene. J Biol Chem 2655:4652–4656

    Google Scholar 

  • Mallonee DH, Glatz BA, Pattee PA (1982) Chromosomal mapping of a gene affecting enterotoxin A production in Staphylococcus aureus. Appl Environ Microbiol 43:397–402

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbour Laboratory Press, Cold Spring Habour, New York

    Google Scholar 

  • Meighen EA (1988) Enzymes and genes from the lux operons of bioluminescent bacteria. Annu Rev Microbiol 42:151–176

    Google Scholar 

  • Menzel R, Gellert M (1987) Fusions of the Escherichia coli gyrA and gyrB control regions to the galactokinase gene are inducible by coumermycin treatment. J Bacteriol 169:1272–1278

    Google Scholar 

  • Miller JF, Mekalanos JJ, Falkow S (1989) Coordinate regulation and sensory transduction in the control of bacterial virulence. Science 243:916–922

    Google Scholar 

  • Morfeldt T, Janzon L, Arvidson S, Lofdahl S (1988) Cloning of a chromosomal locus (exp) which regulates the expression of several exoprotein genes in Staphylococcus aureus. Mol Gen Genet 211:435–440

    Google Scholar 

  • Novick RP (1963) Properties of a cryptic high-frequency transducing phase in Staphylococcus aureus. Virology 33:155–166

    Google Scholar 

  • Oskouian B, Stewart GC (1990) Repression and catabolite repression of the lactose operon of Staphylococcus aureus. J Bacteriol 172:3804–3812

    Google Scholar 

  • Peng HI, Novick RP, Kreiswirth B, Kornblum J, Schlievert P (1988) Cloning, characterisation and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus. J Bacteriol 170:4365–4372

    Google Scholar 

  • Ray C, Hay RE, Carter HL, Moran CP (1985) Mutations that affect utilisation of a promoter in stationary-phase Bacillus subtilis. J Bacteriol 163:610–614

    Google Scholar 

  • Rescei P, Kreiswirth B, O'Reilly M, Schlievert P, Gruss A, Novick RP (1986) Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Mol Gen Genet 202:58–61

    Google Scholar 

  • Ronson CW, Nixon BT, Ausubel FM (1987) Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell 49:579–581

    Google Scholar 

  • Sakuri S, Suzuki H, Kondo I (1988) DNA sequencing of the eta gene coding for staphylococcal exfoliative toxin serotype A. J Gen Microbiol 134:711–717

    Google Scholar 

  • Stock JB, Ninfa AJ, Stock AM (1989) Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490

    Google Scholar 

  • Stock JB, Stock AM, Mottonen J (1990) Signal transduction in bacteria. Nature 344:395–400

    Google Scholar 

  • Sutcliffe JA, Gootz TD, Barrett JF (1989) Biochemical characteristics and physiological significance of major DNA topoisomerases. Antimicrob Agents Chemother 33:2027–2033

    Google Scholar 

  • O'Toole P, Foster TJ (1986) Molecular cloning and expression of the epidermolytic toxin A gene of Staphylococcus aureus. Microbial Pathogenesis 1:583–594

    Google Scholar 

  • O'Toole P, Foster TJ (1987) Nucleotide sequence of the epidermolytic toxin A gene of Staphylococcus aureus. J Bacteriol 169:3910–3915

    Google Scholar 

  • Wilson C, Skinner S, Shaw W (1981) Analysis of two chloramphenicol resistance plasmids from Staphylococcus aureus: Insertional inactivation of chloramphenicol resistance, mapping of restriction sites, and construction of cloning vehicles. Plasmid 5:245–258

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1983) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC vectors. Gene 33:103–119

    Google Scholar 

  • Zukowski MM, Gaffney DF, Speck D, Kauffmann M, Findeli A, Wisecup A, Lecocq J (1983) Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene. Proc Natl Acad Sci USA 80:1101–1105

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, Moyne Institute, Trinity College, Dublin 2, Republic of Ireland

    Brian J. Sheehan & Timothy J. Foster

  2. Molecular Genetics Laboratory, Department of Biochemistry, University of Dundee, DD1 4HN, Dundee, Scotland, UK

    Charles J. Dorman

  3. Department of Microbiology, A.F.R.C. Institute of Food Research, Reading Laboratory, RG2, 9AT, Shinfield, Reading, UK

    Simon Park

  4. Department of Applied Biochemistry and Food Science, School of Agriculture, University of Nottingham, LE12 5RD, Sutton Bonington, Loughborough, UK

    Gordon S. A. B. Stewart

Authors
  1. Brian J. Sheehan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Timothy J. Foster
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Charles J. Dorman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Simon Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gordon S. A. B. Stewart
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by W. Goebel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sheehan, B.J., Foster, T.J., Dorman, C.J. et al. Osmotic and growth-phase dependent regulation of the eta gene of Staphylococcus aureus: a role for DNA supercoiling. Molec. Gen. Genet. 232, 49–57 (1992). https://doi.org/10.1007/BF00299136

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00299136

Key words

Search

Navigation