Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Jul;172(7):3661–3668. doi: 10.1128/jb.172.7.3661-3668.1990

Identification of cis-acting sequences required for translational autoregulation of the ermC methylase.

F Breidt 1, D Dubnau 1
PMCID: PMC213340  PMID: 2113909

Abstract

ermC methylase gene expression has been shown to be limited by translational autorepression, presumably due to methylase binding to ermC mRNA. It was found that this repression occurs in trans, yielding a 50% reduction in translation of an ermC-lacZ fusion mRNA. We investigated the ermC mRNA sequences required for translational repression in vivo. A series of deletions identified sequences in the 5' regulatory region that were required for translational repression. These included sequences of the 5' stem-loop structure that were not required for induction, as well as some that were required. The implications of these results for regulation are discussed.

Full text

PDF
3661

Images in this article

Selected References

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

  1. Albano M., Breitling R., Dubnau D. A. Nucleotide sequence and genetic organization of the Bacillus subtilis comG operon. J Bacteriol. 1989 Oct;171(10):5386–5404. doi: 10.1128/jb.171.10.5386-5404.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albano M., Dubnau D. A. Cloning and characterization of a cluster of linked Bacillus subtilis late competence mutations. J Bacteriol. 1989 Oct;171(10):5376–5385. doi: 10.1128/jb.171.10.5376-5385.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Albano M., Hahn J., Dubnau D. Expression of competence genes in Bacillus subtilis. J Bacteriol. 1987 Jul;169(7):3110–3117. doi: 10.1128/jb.169.7.3110-3117.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bechhofer D. H., Dubnau D. Induced mRNA stability in Bacillus subtilis. Proc Natl Acad Sci U S A. 1987 Jan;84(2):498–502. doi: 10.1073/pnas.84.2.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Denoya C. D., Bechhofer D. H., Dubnau D. Translational autoregulation of ermC 23S rRNA methyltransferase expression in Bacillus subtilis. J Bacteriol. 1986 Dec;168(3):1133–1141. doi: 10.1128/jb.168.3.1133-1141.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denoya C. D., Dubnau D. Site and substrate specificity of the ermC 23S rRNA methyltransferase. J Bacteriol. 1987 Aug;169(8):3857–3860. doi: 10.1128/jb.169.8.3857-3860.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Denoya C., Dubnau D. Mono- and dimethylating activities and kinetic studies of the ermC 23 S rRNA methyltransferase. J Biol Chem. 1989 Feb 15;264(5):2615–2624. [PubMed] [Google Scholar]
  9. Dubnau D. Induction of ermC requires translation of the leader peptide. EMBO J. 1985 Feb;4(2):533–537. doi: 10.1002/j.1460-2075.1985.tb03661.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gryczan T. J., Grandi G., Hahn J., Grandi R., Dubnau D. Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance. Nucleic Acids Res. 1980 Dec 20;8(24):6081–6097. doi: 10.1093/nar/8.24.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hahn J., Grandi G., Gryczan T. J., Dubnau D. Translational attenuation of ermC: a deletion analysis. Mol Gen Genet. 1982;186(2):204–216. doi: 10.1007/BF00331851. [DOI] [PubMed] [Google Scholar]
  12. Horinouchi S., Weisblum B. Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7079–7083. doi: 10.1073/pnas.77.12.7079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iordănescu S. Three distinct plasmids originating in the same Staphylococcus aureus strain. Arch Roum Pathol Exp Microbiol. 1976 Jan-Jun;35(1-2):111–118. [PubMed] [Google Scholar]
  14. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lai C. J., Dahlberg J. E., Weisblum B. Structure of an inducibly methylatable nucleotide sequence in 23S ribosomal ribonucleic acid from erythromycin-resistant Staphylococcus aureus. Biochemistry. 1973 Jan 30;12(3):457–460. doi: 10.1021/bi00727a015. [DOI] [PubMed] [Google Scholar]
  16. Lai C. J., Weisblum B., Fahnestock S. R., Nomura M. Alteration of 23 S ribosomal RNA and erythromycin-induced resistance to lincomycin and spiramycin in Staphylococcus aureus. J Mol Biol. 1973 Feb 15;74(1):67–72. doi: 10.1016/0022-2836(73)90355-0. [DOI] [PubMed] [Google Scholar]
  17. Mahler I., Halvorson H. O. Two erythromycin-resistance plasmids of diverse origin and their effect on sporulation in Bacillus subtilis. J Gen Microbiol. 1980 Sep;120(1):259–263. doi: 10.1099/00221287-120-1-259. [DOI] [PubMed] [Google Scholar]
  18. Mayford M., Weisblum B. Messenger RNA from Staphylococcus aureus that specifies macrolide-lincosamide-streptogramin resistance. Demonstration of its conformations and of the leader peptide it encodes. J Mol Biol. 1985 Oct 20;185(4):769–780. doi: 10.1016/0022-2836(85)90061-0. [DOI] [PubMed] [Google Scholar]
  19. Monod M., Denoya C., Dubnau D. Sequence and properties of pIM13, a macrolide-lincosamide-streptogramin B resistance plasmid from Bacillus subtilis. J Bacteriol. 1986 Jul;167(1):138–147. doi: 10.1128/jb.167.1.138-147.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Narayanan C. S., Dubnau D. An in vitro study of the translational attenuation model of ermC regulation. J Biol Chem. 1987 Feb 5;262(4):1756–1765. [PubMed] [Google Scholar]
  21. Narayanan C. S., Dubnau D. Demonstration of erythromycin-dependent stalling of ribosomes on the ermC leader transcript. J Biol Chem. 1987 Feb 5;262(4):1766–1771. [PubMed] [Google Scholar]
  22. Narayanan C. S., Dubnau D. Evidence for the translational attenuation model: ribosome-binding studies and structural analysis with an in vitro run-off transcript of ermC. Nucleic Acids Res. 1985 Oct 25;13(20):7307–7326. doi: 10.1093/nar/13.20.7307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nomura M., Gourse R., Baughman G. Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem. 1984;53:75–117. doi: 10.1146/annurev.bi.53.070184.000451. [DOI] [PubMed] [Google Scholar]
  24. Reeve J. N., Mendelson N. H., Coyne S. I., Hallock L. L., Cole R. M. Minicells of Bacillus subtilis. J Bacteriol. 1973 May;114(2):860–873. doi: 10.1128/jb.114.2.860-873.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shivakumar A. G., Dubnau D. Characterization of a plasmid-specified ribosome methylase associated with macrolide resistance. Nucleic Acids Res. 1981 Jun 11;9(11):2549–2562. doi: 10.1093/nar/9.11.2549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shivakumar A. G., Hahn J., Dubnau D. Studies on the synthesis of plasmid-coded proteins and their control in Bacillus subtilis minicells. Plasmid. 1979 Apr;2(2):279–289. doi: 10.1016/0147-619x(79)90046-5. [DOI] [PubMed] [Google Scholar]
  27. Shivakumar A. G., Hahn J., Grandi G., Kozlov Y., Dubnau D. Posttranscriptional regulation of an erythromycin resistance protein specified by plasmic pE194. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3903–3907. doi: 10.1073/pnas.77.7.3903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weisblum B., Graham M. Y., Gryczan T., Dubnau D. Plasmid copy number control: isolation and characterization of high-copy-number mutants of plasmid pE194. J Bacteriol. 1979 Jan;137(1):635–643. doi: 10.1128/jb.137.1.635-643.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES