Sequences involved in growth-phase-dependent expression and glucose repression of a α-amylase gene Free

Abstract

In glycerol-grown, but not in glucose-grown cultures, expression in TK24 of the cloned α-amylase gene () of is switched on toward the end of exponential growth. During this period, expression is further inducible by maltotriose. We showed that a 378 bp fragment, extending from position −204 through to + 174 (relative to the transcriptional start site), included -acting sequences involved in regulation. When this fragment was present on a multicopy plasmid, the growth-phase-dependent expression conferred by a DNA fragment cloned on a compatible low-copy-number plasmid was greatly enhanced, as if negative regulators were being titrated. A study of the regulation of expression in variants with deletions in the promoter region indicated that a direct repeat (DR) between positions −124 and −106 (relative to the transcriptional start site) and an inverted repeat (IR) between positions +9 and +24 were good candidates for secondary and primary operator sites, respectively. Deletion of a 29 bp fragment containing the IR rendered expression partly growth-phase-independent, resistant to glucose repression, and insensitive to maltotriose induction.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-5-1059
1994-05-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/5/mic-140-5-1059.html?itemId=/content/journal/micro/10.1099/13500872-140-5-1059&mimeType=html&fmt=ahah

References

  1. Adamidis T., Champness W. 1992; Genetic analysis of abs B, a Streptomjces coelicolor locus involved in global antibiotic regulation. J Bacterial 174:4622–4628
    [Google Scholar]
  2. Adamidis T., Riggle P., Champness W. 1990; Mutations in a new Streptomjces coelicolor locus which globally block antibiotic biosynthesis but not sporulation. J Bacterial 172:2962–2969
    [Google Scholar]
  3. Angell S., Schwarz E., Bibb M. J. 1992; The glucose kinase gene of Streptomjces coelicolor A3(2): its nucleotide sequence, transcriptional analysis and role in glucose repression. Mol Microbiol 6:2833–2844
    [Google Scholar]
  4. Boccard F., Smokvina T., Pernodet J.-L., Friedmann A., Guerineau M. 1989; Structural analysis of loci involved in pSAM2 site-specific integration in Streptomjces.. Plasmid 21:59–70
    [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
    [Google Scholar]
  6. Botsford J.L., Harman J. G. 1992; Cyclic AMP in prokaryotes. Microbiol Rep 56:100–122
    [Google Scholar]
  7. Buttner M. J., Fearnley I. M., Bibb M. J. 1987; The agarase gene (dag A) of Streptomjces coelicolor A3(2): nucleotide sequence and transcriptional analysis. Mol and Gen Genet 209:101–109
    [Google Scholar]
  8. Ceska M. 1971; Enzymatic catalysis in solidified media. Eur J Biochem 22:186–192
    [Google Scholar]
  9. Chambliss G.H. 1993; Carbon source-mediated catabolite repression.. In Bacillus subtilis and Other Gram-Positive Bacteria: Bio-chemistry, Physiology, and Molecular Genetics pp. 213–219 Edited by Sonenshein A. L., Hoch J., Losick R. . Washington, DC: American Society for Microbiology;
    [Google Scholar]
  10. Collado-Vides J, Magasanik B., Gratia J. D. 1991; Control site location and transcriptional regulation in Escherichia coli.. Microbiol Rev 55:371–394
    [Google Scholar]
  11. Daza A., Gill J. A., Vigal T., Martin J. F. 1990; Cloning and characterization of a gene from Streptomjces griseus that increases production of extracellular enzymes in several species of Streptomjces.. Mol and Gen Genet 222:384–392
    [Google Scholar]
  12. Daza A., Martin J. F., Vigal T., Gil J. A. 1991; Analysis of the promoter region of saf, a Streptomjces griseus gene that increases production of extracellular enzymes. Gene 108:63–71
    [Google Scholar]
  13. Delic I., Robbins P., Westpheling J. 1992; Direct repeat sequences are implicated in the regulation of two Streptomjces chitinase promoters that are subject to carbon catabolite control. Proc Natl Acad Sci USA 89:1885–1889
    [Google Scholar]
  14. Fernandez-Moreno M.A., Martin-Triana A. J., Martinez E., Niemi H., Kieser H. M., Hopwood D. A., Malpartida F. 1992; aba A, a new pleiotropic regulatory locus for antibiotic production in Streptomjces coelicolor.. J Bacteriol 174:2958–2967
    [Google Scholar]
  15. Fisher S.H., Sonenshein A. L. 1991; Control of carbon and nitrogen metabolism in Bacillus subtilis.. Annu Rev Microbiol 45:107–135
    [Google Scholar]
  16. Fornwald J. A., Schmidt F. J., Adams C. W., Rosenberg M., Brawner M. E. 1987; Two promoters, one inducible and one constitutive, control transcription of the Streptomjces lividans galactose operon. Proc Natl Acad Sci USA 84:2130–2134
    [Google Scholar]
  17. Gentz R., Langner A., Chang A. C. Y., Cohen S. N., Bujard H. 1981; Cloning and analysis of strong promoters is made possible by the downstream placement of an RNA termination signal. Proc Natl Acad Sci USA 78:4936–4940
    [Google Scholar]
  18. Gramajo H. C., Takano E., Bibb M. J. 1993; Stationary-phase production of the antibiotic actinorhodin in Streptomjces coelicolor A3(2) is transcriptionally regulated. Mol Microbiol 7:837–845
    [Google Scholar]
  19. Herbert D., Phipps P. J., Strange R. E. 1971; Chemical analysis of microbial cells.. In Methods in Microbiology pp. 244–249 Edited by Norris J. R., Ribbons D. W. . New York: Academic Press;
    [Google Scholar]
  20. Hopwood D. A., Kieser T., Wright H. M., Bibb M. J. 1983; Plasmids, recombination and chromosome mapping in Streptomjces lividans 66. J Gen Microbiol 129:2257–2269
    [Google Scholar]
  21. Hopwood D. A., Bibb M. J., Chater K. F., Kieser T., Bruton C. J., Kieser H. M., Lydiate D. J., Smith C. P., Ward J. M., Schrempf H. 1985; Genetic Manipulation of Streptomjces: a laboratory Manual.. Norwich, UK: John Innes Foundation;
    [Google Scholar]
  22. Horinouchi S., Beppu T. 1992; Regulation of secondary metabolism and cell differentiation in Streptomjces: A-factor as a microbial hormone and the Afs R protein as a component of a two-component regulatory system. Gene 115:167–172
    [Google Scholar]
  23. Ishizuka H., Horinouchi S., Kieser H.M., Hopwood D. A., Beppu T. 1992; A putative two-component regulatory system involved in secondary metabolism in Streptomjces spp. J Bacteriol 174:7585–7594
    [Google Scholar]
  24. Klein B., Foreman J. A., Searcy R. L. 1969; The synthesis and utilisation of Cibacron-blue-amylose: a new chromogenic substrate for determination of amylase activity. Anal Biochem 31:412–425
    [Google Scholar]
  25. Lydiate D. J., Malpartida F., Hopwood D. A. 1985; The Streptomjces plasmid SCP2*: its functional analysis and development into useful cloning vectors. Gene 35:223–235
    [Google Scholar]
  26. Mattern S. G., Brawner M. E., Westpheling J. 1993; Identification of a complex operator for gal P1, the glucose-sensitive, galactose-dependent promoter of the Streptomjces galactose operon. J Bacteriol 175:1213–1220
    [Google Scholar]
  27. Matthews K.S. 1992; DNA looping. Microbiol Rev 56:123–136
    [Google Scholar]
  28. Oskouian B., Stewart G. C. 1990; Repression and catabolite repression of the lactose operon of Staphylococcus aureus.. J Bacteriol 172:3804–3812
    [Google Scholar]
  29. Postma P. W., Lengeler J. W., Jacobson G. R. 1993; Phospho- enolpyruvate: carbohydrate phosphotransferase system of bacteria. Microbiol Rep 57:543–594
    [Google Scholar]
  30. Prentki P., Krisch H. M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313
    [Google Scholar]
  31. Rygus T., Hillen W. 1992; Catabolite repression of the xyl operon in Bacillus megaterium.. J Bacterial 17:3049–3055
    [Google Scholar]
  32. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  33. Sizemore C, Wieland B., Gotz F., Hillen W. 1992; Regulation of Staphylococcus xylosus xylose utilization genes at the molecular level. J Bacteriol 174:3042–3048
    [Google Scholar]
  34. Smith C.P., Chater K. F. 1988; Structure and regulation of controlling sequences for the Streptomyces coelicolor glycerol operon. J Mol Biol 204:569–580
    [Google Scholar]
  35. Smokvina T., Mazodier P., Boccard F., Thompson C. J., Guerineau M. 1990; Construction of a series of pSAM2-based integrative vectors for use in actinomycetes. Gene 94:53–59
    [Google Scholar]
  36. Stewart G.C. 1993; Catabolite repression in the gram-positive bacteria: generation of negative regulators of transcription. J Cell Biochem 51:25–28
    [Google Scholar]
  37. Stock J. B., Ninfa A. J., Stock A. M. 1989; Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490
    [Google Scholar]
  38. Stutzman-Engwall K.J., Otten S. L., Hutchinson C. R. 1992; Regulation of secondary metabolism in Streptomyces spp. and overproduction of daunorubicin in Streptomyces peuceticus.. J Bacteriol 174:144–154
    [Google Scholar]
  39. Takano E., Gramajo H. C., Strauch E., Andres N., White J., Bibb M. J. 1992; Transcriptional regulation of the red D transcriptional activator gene accounts for growth-phase-dependent production of the antibiotic undecylprodigiosin in Streptomyces coelicolor A3(2). Mol Microbiol 6:2797–2804
    [Google Scholar]
  40. Virolle M.J., Bibb M. J. 1988; Cloning, characterization and regulation of an alpha-amylase gene from Streptomyces limosus.. Mol Microbiol 2:197–208
    [Google Scholar]
  41. Virolle M. J., Morris V. J., Bibb M. J. 1990; A simple and reliable turbidimetric and kinetic assay for alpha-amylase that is readily applied to culture supernatants and cell extracts. J Ind Microbiol 5:295–302
    [Google Scholar]
  42. Ward J. M., Janssen G. R., Kieser T., Bibb M. J., Buttner M. J., Bibb M. J. 1986; Construction and characterisation of a series of multi-copy promoter-probe plasmid vectors for Streptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicator.. Mol J Gen Genet 203:468–478
    [Google Scholar]
  43. Wong S. L., Wang L. F., Doi R. H. 1988; Cloning and nucleotide sequence of sen N, a novel ‘Bacillus natto’’ (B. subtilis) gene that regulates expression of extracellular protein genes. J Gen Microbiol 134:3269–3276
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-5-1059
Loading
/content/journal/micro/10.1099/13500872-140-5-1059
Loading

Data & Media loading...

Most cited Most Cited RSS feed