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. 1993 Mar;175(5):1213–1220. doi: 10.1128/jb.175.5.1213-1220.1993

Identification of a complex operator for galP1, the glucose-sensitive, galactose-dependent promoter of the Streptomyces galactose operon.

S G Mattern 1, M E Brawner 1, J Westpheling 1
PMCID: PMC193204  PMID: 7680340

Abstract

The galP1 promoter is responsible for galactose-dependent, glucose-sensitive transcription of the galactose utilization operon of Streptomyces coelicolor and Streptomyces lividans. We describe the characterization of mutations that were positioned directly upstream of the apparent transcription start site of galP1 and that resulted in deregulated expression. Certain combinations of base changes within a series of hexamers that lie within two pairs of direct repeat sequences resulted in significant expression from galP1 in the absence of inducer. These motifs are further implicated in regulation by the observation that DNA fragments containing the hexamers and direct repeat sequences resulted in increased transcription from the chromosomal copy of galP1 on multicopy plasmids in the absence of galactose. We suggest that these hexamers and direct repeat sequences constitute an operator for the negative regulation of the Streptomyces gal operon.

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Selected References

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  1. Adams C. W., Fornwald J. A., Schmidt F. J., Rosenberg M., Brawner M. E. Gene organization and structure of the Streptomyces lividans gal operon. J Bacteriol. 1988 Jan;170(1):203–212. doi: 10.1128/jb.170.1.203-212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adams C. W., Hatfield G. W. Effects of promoter strengths and growth conditions on copy number of transcription-fusion vectors. J Biol Chem. 1984 Jun 25;259(12):7399–7403. [PubMed] [Google Scholar]
  3. Adhya S., Echols H. Glucose effect and the galactose enzymes of Escherichia coli: correlation between glucose inhibition of induction and inducer transport. J Bacteriol. 1966 Sep;92(3):601–608. doi: 10.1128/jb.92.3.601-608.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Adhya S., Miller W. Modulation of the two promoters of the galactose operon of Escherichia coli. Nature. 1979 Jun 7;279(5713):492–494. doi: 10.1038/279492a0. [DOI] [PubMed] [Google Scholar]
  5. BERTANI G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol. 1951 Sep;62(3):293–300. doi: 10.1128/jb.62.3.293-300.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. BUTTIN G. M'ECANISMES R'EGULATEURS DANS LA BIOSYNTH'ESE DES ENZYMES DU M'ETABOLISME DU GALACTOSE CHEZ ESCHERICHIA COLI K12. II. LE D'ETERMINISME G'EN'ETIQUE DE LA R'EGULATION. J Mol Biol. 1963 Aug;7:183–205. doi: 10.1016/s0022-2836(63)80045-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  9. Brunelle A., Schleif R. Determining residue-base interactions between AraC protein and araI DNA. J Mol Biol. 1989 Oct 20;209(4):607–622. doi: 10.1016/0022-2836(89)90598-6. [DOI] [PubMed] [Google Scholar]
  10. Buttner M J, Fearnley I M, Bibb M J. The agarase gene (dagA) of Streptomyces coelicolor A3(2): nucleotide sequence and transcriptional analysis. Mol Gen Genet. 1987 Aug;209(1):101–109. doi: 10.1007/BF00329843. [DOI] [PubMed] [Google Scholar]
  11. Buttner M. J., Smith A. M., Bibb M. J. At least three different RNA polymerase holoenzymes direct transcription of the agarase gene (dagA) of Streptomyces coelicolor A3(2). Cell. 1988 Feb 26;52(4):599–607. doi: 10.1016/0092-8674(88)90472-2. [DOI] [PubMed] [Google Scholar]
  12. Champness W. C. New loci required for Streptomyces coelicolor morphological and physiological differentiation. J Bacteriol. 1988 Mar;170(3):1168–1174. doi: 10.1128/jb.170.3.1168-1174.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Delic I., Robbins P., Westpheling J. Direct repeat sequences are implicated in the regulation of two Streptomyces chitinase promoters that are subject to carbon catabolite control. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1885–1889. doi: 10.1073/pnas.89.5.1885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  16. Fornwald J. A., Schmidt F. J., Adams C. W., Rosenberg M., Brawner M. E. Two promoters, one inducible and one constitutive, control transcription of the Streptomyces lividans galactose operon. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2130–2134. doi: 10.1073/pnas.84.8.2130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hendrickson W., Schleif R. A dimer of AraC protein contacts three adjacent major groove regions of the araI DNA site. Proc Natl Acad Sci U S A. 1985 May;82(10):3129–3133. doi: 10.1073/pnas.82.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ho Y. S., Pfarr D., Strickler J., Rosenberg M. Characterization of the transcription activator protein C1 of bacteriophage P22. J Biol Chem. 1992 Jul 15;267(20):14388–14397. [PubMed] [Google Scholar]
  19. Ho Y. S., Wulff D. L., Rosenberg M. Bacteriophage lambda protein cII binds promoters on the opposite face of the DNA helix from RNA polymerase. Nature. 1983 Aug 25;304(5928):703–708. doi: 10.1038/304703a0. [DOI] [PubMed] [Google Scholar]
  20. Ikeda H., Seno E. T., Bruton C. J., Chater K. F. Genetic mapping, cloning and physiological aspects of the glucose kinase gene of Streptomyces coelicolor. Mol Gen Genet. 1984;196(3):501–507. doi: 10.1007/BF00436199. [DOI] [PubMed] [Google Scholar]
  21. Ingram C., Brawner M., Youngman P., Westpheling J. xylE functions as an efficient reporter gene in Streptomyces spp.: use for the study of galP1, a catabolite-controlled promoter. J Bacteriol. 1989 Dec;171(12):6617–6624. doi: 10.1128/jb.171.12.6617-6624.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Irani M. H., Orosz L., Adhya S. A control element within a structural gene: the gal operon of Escherichia coli. Cell. 1983 Mar;32(3):783–788. doi: 10.1016/0092-8674(83)90064-8. [DOI] [PubMed] [Google Scholar]
  23. Kieser T., Hopwood D. A., Wright H. M., Thompson C. J. pIJ101, a multi-copy broad host-range Streptomyces plasmid: functional analysis and development of DNA cloning vectors. Mol Gen Genet. 1982;185(2):223–228. doi: 10.1007/BF00330791. [DOI] [PubMed] [Google Scholar]
  24. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  25. Lomovskaya N. D., Mkrtumian N. M., Gostimskaya N. L., Danilenko V. N. Characterization of temperate actinophage phi C31 isolated from Streptomyces coelicolor A3(2). J Virol. 1972 Feb;9(2):258–262. doi: 10.1128/jvi.9.2.258-262.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
  27. Musso R. E., Di Lauro R., Adhya S., de Crombrugghe B. Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell. 1977 Nov;12(3):847–854. doi: 10.1016/0092-8674(77)90283-5. [DOI] [PubMed] [Google Scholar]
  28. Robbins P. W., Overbye K., Albright C., Benfield B., Pero J. Cloning and high-level expression of chitinase-encoding gene of Streptomyces plicatus. Gene. 1992 Feb 1;111(1):69–76. doi: 10.1016/0378-1119(92)90604-n. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Seno E. T., Bruton C. J., Chater K. F. The glycerol utilization operon of Streptomyces coelicolor: genetic mapping of gyl mutations and the analysis of cloned gylDNA. Mol Gen Genet. 1984;193(1):119–128. doi: 10.1007/BF00327424. [DOI] [PubMed] [Google Scholar]
  31. Shapiro J. A., Adhya S. L. The galactose operon of E. coli K-12. II. A deletion analysis of operon structure and polarity. Genetics. 1969 Jun;62(2):249–264. doi: 10.1093/genetics/62.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smith C. P., Chater K. F. Cloning and transcription analysis of the entire glycerol utilization (gylABX) operon of Streptomyces coelicolor A3(2) and identification of a closely associated transcription unit. Mol Gen Genet. 1988 Jan;211(1):129–137. doi: 10.1007/BF00338403. [DOI] [PubMed] [Google Scholar]
  33. Smith C. P., Chater K. F. Structure and regulation of controlling sequences for the Streptomyces coelicolor glycerol operon. J Mol Biol. 1988 Dec 5;204(3):569–580. doi: 10.1016/0022-2836(88)90356-7. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Thompson C. J., Ward J. M., Hopwood D. A. Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol. 1982 Aug;151(2):668–677. doi: 10.1128/jb.151.2.668-677.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vidal-Ingigliardi D., Richet E., Raibaud O. Two MalT binding sites in direct repeat. A structural motif involved in the activation of all the promoters of the maltose regulons in Escherichia coli and Klebsiella pneumoniae. J Mol Biol. 1991 Mar 20;218(2):323–334. doi: 10.1016/0022-2836(91)90715-i. [DOI] [PubMed] [Google Scholar]
  37. Virolle M. J., Bibb M. J. Cloning, characterization and regulation of an alpha-amylase gene from Streptomyces limosus. Mol Microbiol. 1988 Mar;2(2):197–208. doi: 10.1111/j.1365-2958.1988.tb00021.x. [DOI] [PubMed] [Google Scholar]
  38. Westpheling J., Brawner M. Two transcribing activities are involved in expression of the Streptomyces galactose operon. J Bacteriol. 1989 Mar;171(3):1355–1361. doi: 10.1128/jb.171.3.1355-1361.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. Zukowski M. M., Gaffney D. F., Speck D., Kauffmann M., Findeli A., Wisecup A., Lecocq J. P. Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1101–1105. doi: 10.1073/pnas.80.4.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]

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