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DNA sequence and regulation of ermD, a macrolide-lincosamide-streptogramin B resistance element from Bacillus licheniformis

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Summary

The DNA sequence of ermD, a macrolide-lincosamide-streptogramin B (MLS) resistance determinant cloned from the chromosome of Bacillus licheniformis, has been determined, ermD encodes an erythromycin inducible protein of molecular weight 32,796. S1 nuclease mapping of the ermD promoter has revealed the presence of an approximately 354 base leader sequence on the ermD transcript. This leader contains a short open reading frame sufficient to encode a 14 amino acid peptide, which is preceded by a potential ribosomal binding site. The leader sequence has the potential to fold into several base paired structures, in some of which the ribosomal binding site for the ermD product would be sequestered. Deletion analysis demonstrated that the leader contains regulatory sequences. Removal of the ermD promoter and fusion to an upstream promoter did not interfere with induction, strongly suggestion that ermD regulation is posttranscriptional. Based on these features it appears likely that ermD is regulated by a translational attenuation mechanism, analogous to that suggested for ermC, a resistance element from Staphylococcus aureus (Gryczan et al. 1980; Horinouchi and Weisblum 1980).

Comparison of the ermD sequence and that of its product to two other sequenced MLS determinants reveals substantial phylogenetic relatedness, although the three genes are not homologous by the criterion of Southern blot hybridization.

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References

  • Berk AJ, Sharp PA (1977) Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell 12:721–732

    Google Scholar 

  • Cech TR, Tanner N, Tinoco Jr I, Weir BR, Zuker M, Perlman PS (1983) Secondary structure of the Tetrahymena ribosomal RNA intervening sequence: Structural homology with fungal mitochondrial intervening sequences. Proc Natl Acad Sci USA 80:3903–3907

    Google Scholar 

  • Dayhoff MO, Hunt LT, Hurst-Calderone S (1978a) Composition of proteins. Atlas of Protein Sequence and Structure 5:Suppl 3:363

    Google Scholar 

  • Dayhoff MO, Schwartz RM, Orcutt BC (1978b) A model of evolutionary change in proteins. In: Atlas of protein sequence and structure, 5 (Suppl 3): 345–353. Natl Biomedical Research Foundation, Silver Spring, Md

    Google Scholar 

  • Docherty A, Grandi G, Grandi R, Gryczan TJ, Shivakumar AG, Dubnau D (1981) Naturally occurring macrolite-lincosamide-streptogramin B resistance in Bacillus licheniformis. J Bacteriol 145:129–137

    Google Scholar 

  • Dubnau D, Grandi G, Grandi R, Gryczan TJ, Hahn J, Kozloff Y, Shivakumar AG (1981) Regulation of plasmid specified MLS-resistance in Bacillus subtilis by conformational alteration of RNA structure. In: Levy SB, Clowes RC, Koenig EL (eds) Molecular biology, pathogenicity, and ecology of bacterial plasmids. Plenum Publishing Corp, New York, pp 157–167

    Google Scholar 

  • Favaloro J, Treisman R, Kamen R (1980) Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol 65:718–749

    Google Scholar 

  • Gryczan TJ, Contente S, Dubnau D (1978) Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol 132:318–329

    Google Scholar 

  • Gryczan TJ, Grandi G, Hahn J, Grandi R, Dubnau D (1980) Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance. Nucleic Acids Res 8:6081–6097

    Google Scholar 

  • Gryczan TJ, Hahn J, Contente S, Dubnau D (1982) Replication and incompatibility properties of the plasmid pE194 in Bacillus subtilis. J Bacteriol 152:722–735

    Google Scholar 

  • Gryczan TJ, Israeli-Reches M, Dubnau D (1984) Induction of macrolide-lincosamide-streptogramin B resistance requires ribosomes able to bind inducer. Mol Gen Genet 194:357–361

    Google Scholar 

  • Hahn J, Grandi G, Gryczan TJ, Dubnau D (1982) Translational attenuation of ermC: A deletion analysis. Mol Gen Genet 186:204–216

    Google Scholar 

  • Horinouchi S, Weisblum B (1980) Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci USA 77:7079–7083

    Google Scholar 

  • Horinouchi S, Weisblum B (1981) The control region for erythromycin resistance: Free energy changes related to induction and mutation to constitutive expression. Mol Gen Genet 182:341–348

    Google Scholar 

  • Horinouchi S, Byeon W Weisblum B (1983) A complex attenuator regulates inducible resistance to macrolides, lincosamides, and streptogramin type B antibiotics in Streptococcus sanguis. J Bacteriol 154:1252–1262

    Google Scholar 

  • Kolter R, Yanofsky C (1982) Attenuation in amino acid biosynthetic operons. Annu Rev Genet 16:113–134

    Google Scholar 

  • Lai CJ, Dahlberg JE, Weisblum B (1973a) Structure of an inducibly methylatable nucleotide sequence in 23S ribosomal ribonucleic acid from erythromycin-resistant Staphylococcus aureus. Biochemistry 12:457–460

    Google Scholar 

  • Lai CJ, Weisblum B (1971) Altered methylation of ribosomal RNA in an erythromycin-resistant strain of Staphylococcus aureus. Proc Natl Acad Sci USA 68:856–860

    Google Scholar 

  • Lai CJ, Weisblum B, Fahnestock SR, Nomura M (1973b) Alteration of 23 S ribosomal RNA and erythromycin-induced resistance to lincomycin and spiramycin in Staphylococcus aureus. J Mol Biol 74:67–72

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560

    Google Scholar 

  • Moran CP Jr, Lang N, LeGrice SFJ, Lee G, Stephens M, Sonenshein AL, Pero J, Losick R (1982) Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet 186:339–346

    Google Scholar 

  • Murray CL, Rabinowitz JC (1982a) Species specific translation: Characterization of B. subtilis ribosome binding sites. In: Ganesan AT, Chang S, Hoch JA (eds) Molecular cloning and gene regulation in Bacilli. Academic Press, Inc, New York, pp 271–285

    Google Scholar 

  • Murray CL, Rabinowitz JC (1982b) Nucleotide sequences of transcription and translation initiation regions in Bacillus phage ϕ29 early genes. J Biol Chem 257:1053–1062

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Salser W (1977) Globin mRNA sequences: Analysis of base pairing and evolutionary implications. Cold Spring Harbor Symp Quant Biol 42 (Part 2):985–1002

    Google Scholar 

  • Sanger F, Coulson AR (1978) The use of thin acrylamide gels for DNA sequencing. FEBS Lett 87:107–110

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: Complementary to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Shivakumar AG, Dubnau D (1981) Characterization of a plasmid-specified ribosome methylase associated with macrolide resistance. Nucleic Acid Res 9:2549–2562

    Google Scholar 

  • Shivakumar AG, Hahn J, Dubnau D (1979) Studies on the synthesis of plasmid-coded proteins and their control in Bacillus subtilis minicells. Plasmid 2:279–289

    Google Scholar 

  • Shivakumar AG, Hahn J, Grandi G, Kozlov Y, Dubnau D (1980) Posttranscriptional regulation of an erythromycin resistance protein specified by plasmid pE194. Proc Natl Acad Sci USA 77:3903–3907

    Google Scholar 

  • Skinner RH, Cundliffe E (1982) Dimethylation of adenine and the resistance of Streptomyces crythraeus to erythromycin. J Gen Microbiol 128:2411–2416

    Google Scholar 

  • Smith DR, Calvo JM (1980) Nucleotide sequence of the E. coli gene coding for dihydrofolate reductase. Nucl Acids Res 8:2255–2274

    Google Scholar 

  • Stalker DM, Kolter R, Helinski DR (1979) Nucleotide sequence of the region of an origin of replication of the antibiotic resistance plasmid R6K. Proc Natl Acad Sci USA 76:1150–1154

    Google Scholar 

  • Sutcliffe JG (1978) pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucl Acids Res 5:2721–2728

    Google Scholar 

  • Tu C-PD, Cohen SN (1980) 3′-end labeling of DNA with [α-32P] cordycepin-5′-triphosphate. Gene 10:177–183

    Google Scholar 

  • Weisblum B, Graham MY, Gryczan T, Dubnau D (1979a) Plasmid copy number control: Isolation and characterization of high-copy-number mutants of plasmid pE194. J Bacteriol 137:635–643

    Google Scholar 

  • Weisblum B, Holder SB, Halling SM (1979b) Deoxyribonucleic acid sequence common to staphylococcal and streptococcal plasmids which specify erythromycin resistance. J Bacteriol 138:990–998

    Google Scholar 

  • Zuker M, Stiegler P (1981) Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucl Acids Res 1:133–148

    Google Scholar 

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Communicated by A. Bukhari

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Gryczan, T., Israeli-Reches, M., Del Bue, M. et al. DNA sequence and regulation of ermD, a macrolide-lincosamide-streptogramin B resistance element from Bacillus licheniformis . Molec. Gen. Genet. 194, 349–356 (1984). https://doi.org/10.1007/BF00425543

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  • DOI: https://doi.org/10.1007/BF00425543

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