Abstract
The two genes encoding the class IIS restriction-modification system MboII from Moraxella bovis were cloned separately in two compatible plasmids and expressed in E. coli RR1 delta M15. The nucleotide sequences of the MboII endonuclease (R.MboII) and methylase (M.MboII) genes were determined and the putative start codon of R.MboII was confirmed by amino acid sequence analysis. The mboIIR gene specifies a protein of 416 amino acids (MW: 48,617) while the mboIIM gene codes for a putative 260-residue polypeptide (MW: 30,077). Both genes are aligned in the same orientation. The coding region of the methylase gene ends 11 bp upstream of the start codon of the restrictase gene. Comparing the amino acid sequence of M.MboII with sequences of other N6-adenine methyltransferases reveals a significant homology to M.RsrI, M.HinfI and M.DpnA. Furthermore, M.MboII shows homology to the N4-cytosine methyltransferase BamHI.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bolton B. J., Schmitz G. G., Jarsch M., Comer M. J., Kessler C. Ksp632I, a novel class-IIS restriction endonuclease from Kluyvera sp. strain 632 with the asymmetric hexanucleotide recognition sequence: 5'-CTCTTC(N)1-3' 3'-GAGAAG(N)4-5'. Gene. 1988 Jun 15;66(1):31–43. doi: 10.1016/0378-1119(88)90222-3. [DOI] [PubMed] [Google Scholar]
- Brown N. L., Hutchison C. A., 3rd, Smith M. The specific non-symmetrical sequence recognized by restriction endonuclease MboII. J Mol Biol. 1980 Jun 15;140(1):143–148. doi: 10.1016/0022-2836(80)90360-5. [DOI] [PubMed] [Google Scholar]
- Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ehrlich M., Gama-Sosa M. A., Carreira L. H., Ljungdahl L. G., Kuo K. C., Gehrke C. W. DNA methylation in thermophilic bacteria: N4-methylcytosine, 5-methylcytosine, and N6-methyladenine. Nucleic Acids Res. 1985 Feb 25;13(4):1399–1412. doi: 10.1093/nar/13.4.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ehrlich M., Wilson G. G., Kuo K. C., Gehrke C. W. N4-methylcytosine as a minor base in bacterial DNA. J Bacteriol. 1987 Mar;169(3):939–943. doi: 10.1128/jb.169.3.939-943.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heitman J., Model P. Site-specific methylases induce the SOS DNA repair response in Escherichia coli. J Bacteriol. 1987 Jul;169(7):3243–3250. doi: 10.1128/jb.169.7.3243-3250.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Janulaitis A., Povilionis P., Sasnauskas K. Cloning of the modification methylase gene of Bacillus centrosporus in Escherichia coli. Gene. 1982 Dec;20(2):197–204. doi: 10.1016/0378-1119(82)90038-5. [DOI] [PubMed] [Google Scholar]
- Jen-Jacobson L., Kurpiewski M., Lesser D., Grable J., Boyer H. W., Rosenberg J. M., Greene P. J. Coordinate ion pair formation between EcoRI endonuclease and DNA. J Biol Chem. 1983 Dec 10;258(23):14638–14646. [PubMed] [Google Scholar]
- Kaszubska W., Aiken C., O'Connor C. D., Gumport R. I. Purification, cloning and sequence analysis of RsrI DNA methyltransferase: lack of homology between two enzymes, RsrI and EcoRI, that methylate the same nucleotide in identical recognition sequences. Nucleic Acids Res. 1989 Dec 25;17(24):10403–10425. doi: 10.1093/nar/17.24.10403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kessler C., Manta V. Specificity of restriction endonucleases and DNA modification methyltransferases a review (Edition 3). Gene. 1990 Aug 16;92(1-2):1–248. doi: 10.1016/0378-1119(90)90486-b. [DOI] [PubMed] [Google Scholar]
- Kiss A., Baldauf F. Molecular cloning and expression in Escherichia coli of two modification methylase genes of Bacillus subtilis. Gene. 1983 Jan-Feb;21(1-2):111–119. doi: 10.1016/0378-1119(83)90153-1. [DOI] [PubMed] [Google Scholar]
- Kiss A., Posfai G., Keller C. C., Venetianer P., Roberts R. J. Nucleotide sequence of the BsuRI restriction-modification system. Nucleic Acids Res. 1985 Sep 25;13(18):6403–6421. doi: 10.1093/nar/13.18.6403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kleid D., Humayun Z., Jeffrey A., Ptashne M. Novel properties of a restriction endonuclease isolated from Haemophilus parahaemolyticus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):293–297. doi: 10.1073/pnas.73.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klimasauskas S., Timinskas A., Menkevicius S., Butkienè D., Butkus V., Janulaitis A. Sequence motifs characteristic of DNA[cytosine-N4]methyltransferases: similarity to adenine and cytosine-C5 DNA-methylases. Nucleic Acids Res. 1989 Dec 11;17(23):9823–9832. doi: 10.1093/nar/17.23.9823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Labbé D., Höltke H. J., Lau P. C. Cloning and characterization of two tandemly arranged DNA methyltransferase genes of Neisseria lactamica: an adenine-specific M.NlaIII and a cytosine-type methylase. Mol Gen Genet. 1990 Oct;224(1):101–110. doi: 10.1007/BF00259456. [DOI] [PubMed] [Google Scholar]
- Lauster R. Evolution of type II DNA methyltransferases. A gene duplication model. J Mol Biol. 1989 Mar 20;206(2):313–321. doi: 10.1016/0022-2836(89)90481-6. [DOI] [PubMed] [Google Scholar]
- Looney M. C., Moran L. S., Jack W. E., Feehery G. R., Benner J. S., Slatko B. E., Wilson G. G. Nucleotide sequence of the FokI restriction-modification system: separate strand-specificity domains in the methyltransferase. Gene. 1989 Aug 15;80(2):193–208. doi: 10.1016/0378-1119(89)90284-9. [DOI] [PubMed] [Google Scholar]
- Mannarelli B. M., Balganesh T. S., Greenberg B., Springhorn S. S., Lacks S. A. Nucleotide sequence of the Dpn II DNA methylase gene of Streptococcus pneumoniae and its relationship to the dam gene of Escherichia coli. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4468–4472. doi: 10.1073/pnas.82.13.4468. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClarin J. A., Frederick C. A., Wang B. C., Greene P., Boyer H. W., Grable J., Rosenberg J. M. Structure of the DNA-Eco RI endonuclease recognition complex at 3 A resolution. Science. 1986 Dec 19;234(4783):1526–1541. doi: 10.1126/science.3024321. [DOI] [PubMed] [Google Scholar]
- McClelland M., Nelson M., Cantor C. R. Purification of Mbo II methylase (GAAGmA) from Moraxella bovis: site specific cleavage of DNA at nine and ten base pair sequences. Nucleic Acids Res. 1985 Oct 25;13(20):7171–7182. doi: 10.1093/nar/13.20.7171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClelland M., Nelson M. The effect of site-specific DNA methylation on restriction endonucleases and DNA modification methyltransferases--a review. Gene. 1988 Dec 25;74(1):291–304. doi: 10.1016/0378-1119(88)90305-8. [DOI] [PubMed] [Google Scholar]
- Noyer-Weidner M., Diaz R., Reiners L. Cytosine-specific DNA modification interferes with plasmid establishment in Escherichia coli K12: involvement of rglB. Mol Gen Genet. 1986 Dec;205(3):469–475. doi: 10.1007/BF00338084. [DOI] [PubMed] [Google Scholar]
- Rüther U. pUR 250 allows rapid chemical sequencing of both DNA strands of its inserts. Nucleic Acids Res. 1982 Oct 11;10(19):5765–5772. doi: 10.1093/nar/10.19.5765. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Szybalski W. Universal restriction endonucleases: designing novel cleavage specificities by combining adapter oligodeoxynucleotide and enzyme moieties. Gene. 1985;40(2-3):169–173. doi: 10.1016/0378-1119(85)90039-3. [DOI] [PubMed] [Google Scholar]
- Terry B. J., Jack W. E., Modrich P. Mechanism of specific site location and DNA cleavage by EcoR I endonuclease. Gene Amplif Anal. 1987;5:103–118. [PubMed] [Google Scholar]
- Wharton R. P., Ptashne M. Changing the binding specificity of a repressor by redesigning an alpha-helix. Nature. 1985 Aug 15;316(6029):601–605. doi: 10.1038/316601a0. [DOI] [PubMed] [Google Scholar]
- Wilson G. G. Type II restriction--modification systems. Trends Genet. 1988 Nov;4(11):314–318. doi: 10.1016/0168-9525(88)90109-6. [DOI] [PubMed] [Google Scholar]