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. 1993 Aug;12(8):3183–3191. doi: 10.1002/j.1460-2075.1993.tb05987.x

Autogenous transcriptional activation of a thiostrepton-induced gene in Streptomyces lividans.

D J Holmes 1, J L Caso 1, C J Thompson 1
PMCID: PMC413584  PMID: 7688297

Abstract

Although the antibiotic thiostrepton is best known as an inhibitor of protein synthesis, it also, at extremely low concentrations (< 10(-9) M), induces the expression of a regulon of unknown function in certain Streptomyces species. Here, we report the purification of a Streptomyces lividans thiostrepton-induced transcriptional activator protein, TipAL, whose N-terminus is similar to a family of eubacterial regulatory proteins represented by MerR. TipAL was first purified from induced cultures of S.lividans as a factor which bound to and activated transcription from its own promoter. The tipAL gene was overexpressed in Escherichia coli and TipAL protein purified in a single step using a thiostrepton affinity column. Thiostrepton enhanced binding of TipAL to the promoter and catalysed specific transcription in vitro. TipAS, a second gene product of the same open reading frame consisting of the C-terminal domain of TipAL, is apparently translated using its own in-frame initiation site. Since it is produced in large molar excess relative to TipAL after induction and also binds thiostrepton, it may competitively modulate transcriptional activation.

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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. Amábile-Cuevas C. F., Demple B. Molecular characterization of the soxRS genes of Escherichia coli: two genes control a superoxide stress regulon. Nucleic Acids Res. 1991 Aug 25;19(16):4479–4484. doi: 10.1093/nar/19.16.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ansari A. Z., Chael M. L., O'Halloran T. V. Allosteric underwinding of DNA is a critical step in positive control of transcription by Hg-MerR. Nature. 1992 Jan 2;355(6355):87–89. doi: 10.1038/355087a0. [DOI] [PubMed] [Google Scholar]
  4. Bibb M. J., Cohen S. N. Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet. 1982;187(2):265–277. doi: 10.1007/BF00331128. [DOI] [PubMed] [Google Scholar]
  5. Bläsi U., Nam K., Hartz D., Gold L., Young R. Dual translational initiation sites control function of the lambda S gene. EMBO J. 1989 Nov;8(11):3501–3510. doi: 10.1002/j.1460-2075.1989.tb08515.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Campbell J. L., Richardson C. C., Studier F. W. Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci U S A. 1978 May;75(5):2276–2280. doi: 10.1073/pnas.75.5.2276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collado-Vides J., Magasanik B., Gralla J. D. Control site location and transcriptional regulation in Escherichia coli. Microbiol Rev. 1991 Sep;55(3):371–394. doi: 10.1128/mr.55.3.371-394.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cundliffe E. Mechanism of resistance to thiostrepton in the producing-organism Streptomyces azureus. Nature. 1978 Apr 27;272(5656):792–795. doi: 10.1038/272792a0. [DOI] [PubMed] [Google Scholar]
  10. Cundliffe E. The mode of action of thiostreption in vivo. Biochem Biophys Res Commun. 1971 Aug 20;44(4):912–917. doi: 10.1016/0006-291x(71)90798-4. [DOI] [PubMed] [Google Scholar]
  11. Guilfoile P. G., Hutchinson C. R. A bacterial analog of the mdr gene of mammalian tumor cells is present in Streptomyces peucetius, the producer of daunorubicin and doxorubicin. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8553–8557. doi: 10.1073/pnas.88.19.8553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hall M. N., Gabay J., Débarbouillé M., Schwartz M. A role for mRNA secondary structure in the control of translation initiation. Nature. 1982 Feb 18;295(5850):616–618. doi: 10.1038/295616a0. [DOI] [PubMed] [Google Scholar]
  13. Heltzel A., Lee I. W., Totis P. A., Summers A. O. Activator-dependent preinduction binding of sigma-70 RNA polymerase at the metal-regulated mer promoter. Biochemistry. 1990 Oct 16;29(41):9572–9584. doi: 10.1021/bi00493a011. [DOI] [PubMed] [Google Scholar]
  14. Higgins D. G., Bleasby A. J., Fuchs R. CLUSTAL V: improved software for multiple sequence alignment. Comput Appl Biosci. 1992 Apr;8(2):189–191. doi: 10.1093/bioinformatics/8.2.189. [DOI] [PubMed] [Google Scholar]
  15. Holt T. G., Chang C., Laurent-Winter C., Murakami T., Garrels J. I., Davies J. E., Thompson C. J. Global changes in gene expression related to antibiotic synthesis in Streptomyces hygroscopicus. Mol Microbiol. 1992 Apr;6(8):969–980. doi: 10.1111/j.1365-2958.1992.tb02163.x. [DOI] [PubMed] [Google Scholar]
  16. Kofoid E. C., Parkinson J. S. Tandem translation starts in the cheA locus of Escherichia coli. J Bacteriol. 1991 Mar;173(6):2116–2119. doi: 10.1128/jb.173.6.2116-2119.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Langridge J., Langridge P., Bergquist P. L. Extraction of nucleic acids from agarose gels. Anal Biochem. 1980 Apr;103(2):264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  19. Lokker N. A., Zenke G., Strittmatter U., Fagg B., Movva N. R. Structure-activity relationship study of human interleukin-3: role of the C-terminal region for biological activity. EMBO J. 1991 Aug;10(8):2125–2131. doi: 10.1002/j.1460-2075.1991.tb07746.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Misra T. K., Brown N. L., Fritzinger D. C., Pridmore R. D., Barnes W. M., Haberstroh L., Silver S. Mercuric ion-resistance operons of plasmid R100 and transposon Tn501: the beginning of the operon including the regulatory region and the first two structural genes. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5975–5979. doi: 10.1073/pnas.81.19.5975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murakami T., Holt T. G., Thompson C. J. Thiostrepton-induced gene expression in Streptomyces lividans. J Bacteriol. 1989 Mar;171(3):1459–1466. doi: 10.1128/jb.171.3.1459-1466.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nivinskas R., Vaiskunaite R., Raudonikiene A. An internal AUU codon initiates a smaller peptide encoded by bacteriophage T4 baseplate gene 26. Mol Gen Genet. 1992 Mar;232(2):257–261. doi: 10.1007/BF00280004. [DOI] [PubMed] [Google Scholar]
  24. O'Halloran T. V., Frantz B., Shin M. K., Ralston D. M., Wright J. G. The MerR heavy metal receptor mediates positive activation in a topologically novel transcription complex. Cell. 1989 Jan 13;56(1):119–129. doi: 10.1016/0092-8674(89)90990-2. [DOI] [PubMed] [Google Scholar]
  25. O'Halloran T., Walsh C. Metalloregulatory DNA-binding protein encoded by the merR gene: isolation and characterization. Science. 1987 Jan 9;235(4785):211–214. doi: 10.1126/science.3798107. [DOI] [PubMed] [Google Scholar]
  26. Ogawa H., Imai S., Satoh A., Kojima M. An improved method for the preparation of Streptomycetes and Micromonospora protoplasts. J Antibiot (Tokyo) 1983 Feb;36(2):184–186. doi: 10.7164/antibiotics.36.184. [DOI] [PubMed] [Google Scholar]
  27. Parkhill J., Ansari A. Z., Wright J. G., Brown N. L., O'Halloran T. V. Construction and characterization of a mercury-independent MerR activator (MerRAC): transcriptional activation in the absence of Hg(II) is accompanied by DNA distortion. EMBO J. 1993 Feb;12(2):413–421. doi: 10.1002/j.1460-2075.1993.tb05673.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pestka S., Weiss D., Vince R. Partition of ribosomes in two-polymer aqueous phase systems. Anal Biochem. 1976 Mar;71(1):137–142. doi: 10.1016/0003-2697(76)90020-8. [DOI] [PubMed] [Google Scholar]
  30. Plater R., Robinson J. A. Cloning and sequence of a gene encoding macrotetrolide antibiotic resistance from Streptomyces griseus. Gene. 1992 Mar 1;112(1):117–122. doi: 10.1016/0378-1119(92)90312-d. [DOI] [PubMed] [Google Scholar]
  31. Ryan P. C., Lu M., Draper D. E. Recognition of the highly conserved GTPase center of 23 S ribosomal RNA by ribosomal protein L11 and the antibiotic thiostrepton. J Mol Biol. 1991 Oct 20;221(4):1257–1268. doi: 10.1016/0022-2836(91)90932-v. [DOI] [PubMed] [Google Scholar]
  32. Sadowsky M. J., Cregan P. B., Gottfert M., Sharma A., Gerhold D., Rodriguez-Quinones F., Keyser H. H., Hennecke H., Stacey G. The Bradyrhizobium japonicum nolA gene and its involvement in the genotype-specific nodulation of soybeans. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):637–641. doi: 10.1073/pnas.88.2.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Summers A. O. Untwist and shout: a heavy metal-responsive transcriptional regulator. J Bacteriol. 1992 May;174(10):3097–3101. doi: 10.1128/jb.174.10.3097-3101.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thompson C. J., Skinner R. H., Thompson J., Ward J. M., Hopwood D. A., Cundliffe E. Biochemical characterization of resistance determinants cloned from antibiotic-producing streptomycetes. J Bacteriol. 1982 Aug;151(2):678–685. doi: 10.1128/jb.151.2.678-685.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Thompson J., Cundliffe E. The binding of thiostrepton to 23S ribosomal RNA. Biochimie. 1991 Jul-Aug;73(7-8):1131–1135. doi: 10.1016/0300-9084(91)90156-u. [DOI] [PubMed] [Google Scholar]
  38. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  39. VanBogelen R. A., Neidhardt F. C. Ribosomes as sensors of heat and cold shock in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5589–5593. doi: 10.1073/pnas.87.15.5589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Willey J., Santamaria R., Guijarro J., Geistlich M., Losick R. Extracellular complementation of a developmental mutation implicates a small sporulation protein in aerial mycelium formation by S. coelicolor. Cell. 1991 May 17;65(4):641–650. doi: 10.1016/0092-8674(91)90096-h. [DOI] [PubMed] [Google Scholar]
  41. 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]

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