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Nucleic Acids Res. Mar 11, 1990; 18(5): 1145–1152.
PMCID: PMC330428

Cloning, characterization, and expression in Escherichia coli of the gene coding for the CpG DNA methylase from Spiroplasma sp. strain MQ1(M.SssI).

Abstract

We describe here the cloning, characterization and expression in E. coli of the gene coding for a DNA methylase from Spiroplasma sp. strain MQ1 (M.SssI). This enzyme methylates completely and exclusively CpG sequences. The Spiroplasma gene was transcribed in E. coli using its own promoter. Translation of the entire message required the use of an opal suppressor, suggesting that UGA triplets code for tryptophan in Spiroplasma. Sequence analysis of the gene revealed several UGA triplets, in a 1158 bp long open reading frame. The deduced amino acid sequence revealed in M.SssI all common domains characteristic of bacterial cytosine DNA methylases. The putative sequence recognition domain of M.SssI showed no obvious similarities with that of the mouse DNA methylase, in spite of their common sequence specificity. The cloned enzyme methylated exclusively CpG sequences both in vivo and in vitro. In contrast to the mammalian enzyme which is primarily a maintenance methylase, M.SssI displayed de novo methylase activity, characteristic of prokaryotic cytosine DNA methylases.

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Selected References

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  • Nur I, Szyf M, Razin A, Glaser G, Rottem S, Razin S. Procaryotic and eucaryotic traits of DNA methylation in spiroplasmas (mycoplasmas). J Bacteriol. 1985 Oct;164(1):19–24. [PMC free article] [PubMed]
  • Razin A, Riggs AD. DNA methylation and gene function. Science. 1980 Nov 7;210(4470):604–610. [PubMed]
  • Razin A, Cedar H. DNA methylation in eukaryotic cells. Int Rev Cytol. 1984;92:159–185. [PubMed]
  • Raftery LA, Egan JB, Cline SW, Yarus M. Defined set of cloned termination suppressors: in vivo activity of isogenetic UAG, UAA, and UGA suppressor tRNAs. J Bacteriol. 1984 Jun;158(3):849–859. [PMC free article] [PubMed]
  • Raleigh EA, Wilson G. Escherichia coli K-12 restricts DNA containing 5-methylcytosine. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9070–9074. [PMC free article] [PubMed]
  • Yamao F, Muto A, Kawauchi Y, Iwami M, Iwagami S, Azumi Y, Osawa S. UGA is read as tryptophan in Mycoplasma capricolum. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2306–2309. [PMC free article] [PubMed]
  • Inamine JM, Denny TP, Loechel S, Schaper U, Huang CH, Bott KF, Hu PC. Nucleotide sequence of the P1 attachment-protein gene of Mycoplasma pneumoniae. Gene. 1988 Apr 29;64(2):217–229. [PubMed]
  • Gruenbaum Y, Stein R, Cedar H, Razin A. Methylation of CpG sequences in eukaryotic DNA. FEBS Lett. 1981 Feb 9;124(1):67–71. [PubMed]
  • Wu JC, Santi DV. Kinetic and catalytic mechanism of HhaI methyltransferase. J Biol Chem. 1987 Apr 5;262(10):4778–4786. [PubMed]
  • Lauster R, Trautner TA, Noyer-Weidner M. Cytosine-specific type II DNA methyltransferases. A conserved enzyme core with variable target-recognizing domains. J Mol Biol. 1989 Mar 20;206(2):305–312. [PubMed]
  • Pósfai J, Bhagwat AS, Pósfai G, Roberts RJ. Predictive motifs derived from cytosine methyltransferases. Nucleic Acids Res. 1989 Apr 11;17(7):2421–2435. [PMC free article] [PubMed]
  • Kupper D, Zhou JG, Venetianer P, Kiss A. Cloning and structure of the BepI modification methylase. Nucleic Acids Res. 1989 Feb 11;17(3):1077–1088. [PMC free article] [PubMed]
  • Buhk HJ, Behrens B, Tailor R, Wilke K, Prada JJ, Günthert U, Noyer-Weidner M, Jentsch S, Trautner TA. Restriction and modification in Bacillus subtilis: nucleotide sequence, functional organization and product of the DNA methyltransferase gene of bacteriophage SPR. Gene. 1984 Jul-Aug;29(1-2):51–61. [PubMed]
  • Pósfai G, Baldauf F, Erdei S, Pósfai J, Venetianer P, Kiss A. Structure of the gene coding for the sequence-specific DNA-methyltransferase of the B. subtilis phage SPR. Nucleic Acids Res. 1984 Dec 11;12(23):9039–9049. [PMC free article] [PubMed]
  • Tran-Betcke A, Behrens B, Noyer-Weidner M, Trautner TA. DNA methyltransferase genes of Bacillus subtilis phages: comparison of their nucleotide sequences. Gene. 1986;42(1):89–96. [PubMed]
  • Behrens B, Noyer-Weidner M, Pawlek B, Lauster R, Balganesh TS, Trautner TA. Organization of multispecific DNA methyltransferases encoded by temperate Bacillus subtilis phages. EMBO J. 1987 Apr;6(4):1137–1142. [PMC free article] [PubMed]
  • Wilke K, Rauhut E, Noyer-Weidner M, Lauster R, Pawlek B, Behrens B, Trautner TA. Sequential order of target-recognizing domains in multispecific DNA-methyltransferases. EMBO J. 1988 Aug;7(8):2601–2609. [PMC free article] [PubMed]
  • Trautner TA, Balganesh TS, Pawlek B. Chimeric multispecific DNA methyltransferases with novel combinations of target recognition. Nucleic Acids Res. 1988 Jul 25;16(14A):6649–6658. [PMC free article] [PubMed]
  • Balganesh TS, Reiners L, Lauster R, Noyer-Weidner M, Wilke K, Trautner TA. Construction and use of chimeric SPR/phi 3T DNA methyltransferases in the definition of sequence recognizing enzyme regions. EMBO J. 1987 Nov;6(11):3543–3549. [PMC free article] [PubMed]
  • Pósfai G, Kiss A, Erdei S, Pósfai J, Venetianer P. Structure of the Bacillus sphaericus R modification methylase gene. J Mol Biol. 1983 Nov 5;170(3):597–610. [PubMed]
  • Kiss A, Posfai G, Keller CC, Venetianer P, Roberts RJ. Nucleotide sequence of the BsuRI restriction-modification system. Nucleic Acids Res. 1985 Sep 25;13(18):6403–6421. [PMC free article] [PubMed]
  • Slatko BE, Croft R, Moran LS, Wilson GG. Cloning and analysis of the HaeIII and HaeII methyltransferase genes. Gene. 1988 Dec 25;74(1):45–50. [PubMed]
  • Sullivan KM, Saunders JR. Sequence analysis of the NgoPII methyltransferase gene from Neisseria gonorrhoeae P9: homologies with other enzymes recognizing the sequence 5'-GGCC-3'. Nucleic Acids Res. 1988 May 25;16(10):4369–4387. [PMC free article] [PubMed]
  • Bestor T, Laudano A, Mattaliano R, Ingram V. Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases. J Mol Biol. 1988 Oct 20;203(4):971–983. [PubMed]
  • Caserta M, Zacharias W, Nwankwo D, Wilson GG, Wells RD. Cloning, sequencing, in vivo promoter mapping, and expression in Escherichia coli of the gene for the HhaI methyltransferase. J Biol Chem. 1987 Apr 5;262(10):4770–4777. [PubMed]
  • Karreman C, de Waard A. Cloning and complete nucleotide sequences of the type II restriction-modification genes of Salmonella infantis. J Bacteriol. 1988 Jun;170(6):2527–2532. [PMC free article] [PubMed]
  • Urieli-Shoval S, Gruenbaum Y, Razin A. Sequence and substrate specificity of isolated DNA methylases from Escherichia coli C. J Bacteriol. 1983 Jan;153(1):274–280. [PMC free article] [PubMed]

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