Cloning, Characterization and Heterologous Expression of the SmaI Restriction-modification System

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1989 Dec 11

PMID 2690008

Citations 32

Authors

S Heidmann

W Seifert

C Kessler

H Domdey

Affiliations

Soon will be listed here.

Abstract

The genes coding for the class-II Serratia marcescens restriction-modification system have been cloned and expressed in E. coli. Recombinant clones, restricted incoming phage only poorly; the recombinant plasmids, however, became fully modified in vivo, i.e. completely resistant against digestion with R.SmaI. The determined nucleotide sequence of the cloned system revealed three open reading frames with lengths of 252 bp, 741 bp, and 876 bp. Through various deletion experiments and an insertion-mutation experiment the 876 bp open reading frame could be assigned to the SmaI DNA modification enzyme and the 741 bp open reading frame to the SmaI restriction endonuclease. Mapping of the transcription start sites of the genes revealed that the SmaI endonuclease is transcribed as polycistronic mRNA together with a 252 bp long preceding open reading frame of unknown function. No homology was found when comparing the amino acid sequence of M.SmaI with the published sequences of m5C-specific DNA modification methyltransferases. On the other hand, a stretch of 14 amino acids in the C-proximal region of M.SmaI shows a significant homology to the C-proximal amino acid sequences of the N6A-methyltransferases M.HinfI and M.DpnIIA and the N4C-methyltransferase M.PvuII.

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References

Chandrasegaran S, Lunnen K, Smith H, Wilson G . Cloning and sequencing the HinfI restriction and modification genes. Gene. 1988; 70(2):387-92. DOI: 10.1016/0378-1119(88)90210-7. View

Messer W, Noyer-Weidner M . Timing and targeting: the biological functions of Dam methylation in E. coli. Cell. 1988; 54(6):735-7. DOI: 10.1016/s0092-8674(88)90911-7. View

Posfai J, Bhagwat A, Posfai G, Roberts R . Predictive motifs derived from cytosine methyltransferases. Nucleic Acids Res. 1989; 17(7):2421-35. PMC: 317633. DOI: 10.1093/nar/17.7.2421. View

Wilson G . Cloned restriction-modification systems--a review. Gene. 1988; 74(1):281-9. DOI: 10.1016/0378-1119(88)90304-6. View

McClelland M, Nelson M . The effect of site-specific DNA methylation on restriction endonucleases and DNA modification methyltransferases--a review. Gene. 1988; 74(1):291-304. DOI: 10.1016/0378-1119(88)90305-8. View

Tao T, Walter J, Brennan K, Cotterman M, Blumenthal R . Sequence, internal homology and high-level expression of the gene for a DNA-(cytosine N4)-methyltransferase, M.Pvu II. Nucleic Acids Res. 1989; 17(11):4161-75. PMC: 317926. DOI: 10.1093/nar/17.11.4161. View

Luria S, HUMAN M . A nonhereditary, host-induced variation of bacterial viruses. J Bacteriol. 1952; 64(4):557-69. PMC: 169391. DOI: 10.1128/jb.64.4.557-569.1952. View

Bertani G, Weigle J . Host controlled variation in bacterial viruses. J Bacteriol. 1953; 65(2):113-21. PMC: 169650. DOI: 10.1128/jb.65.2.113-121.1953. View

Linn S, Arber W . Host specificity of DNA produced by Escherichia coli, X. In vitro restriction of phage fd replicative form. Proc Natl Acad Sci U S A. 1968; 59(4):1300-6. PMC: 224867. DOI: 10.1073/pnas.59.4.1300. View

10.

NEEDLEMAN S, Wunsch C . A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970; 48(3):443-53. DOI: 10.1016/0022-2836(70)90057-4. View

11.

Endow S, Roberts R . Two restriction-like enzymes from Xanthomonas malvacearum. J Mol Biol. 1977; 112(3):521-9. DOI: 10.1016/s0022-2836(77)80198-8. View

12.

Gautier F, Bunemann H, Grotjahn L . Analysis of calf-thymus satellite DNA: evidence for specific methylation of cytosine in C-G sequences. Eur J Biochem. 1977; 80(1):175-83. DOI: 10.1111/j.1432-1033.1977.tb11869.x. View

13.

Mann M, Smith H . Specificity of Hpa II and Hae III DNA methylases. Nucleic Acids Res. 1977; 4(12):4211-21. PMC: 343235. DOI: 10.1093/nar/4.12.4211. View

14.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

15.

Grimont P, Grimont F . The genus Serratia. Annu Rev Microbiol. 1978; 32:221-48. DOI: 10.1146/annurev.mi.32.100178.001253. View

16.

Aiba H, Adhya S, de Crombrugghe B . Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981; 256(22):11905-10. View

17.

Youssoufian H, Mulder C . Detection of methylated sequences in eukaryotic DNA with the restriction endonucleases Smai and Xmai. J Mol Biol. 1981; 150(1):133-6. DOI: 10.1016/0022-2836(81)90328-4. View

18.

Hanahan D . Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983; 166(4):557-80. DOI: 10.1016/s0022-2836(83)80284-8. View

19.

McClelland M, Kessler L, Bittner M . Site-specific cleavage of DNA at 8- and 10-base-pair sequences. Proc Natl Acad Sci U S A. 1984; 81(4):983-7. PMC: 344747. DOI: 10.1073/pnas.81.4.983. View

20.

Swinton D, Hattman S, Crain P, Cheng C, Smith D, McCloskey J . Purification and characterization of the unusual deoxynucleoside, alpha-N-(9-beta-D-2'-deoxyribofuranosylpurin-6-yl)glycinamide, specified by the phage Mu modification function. Proc Natl Acad Sci U S A. 1983; 80(24):7400-4. PMC: 389958. DOI: 10.1073/pnas.80.24.7400. View