Home LiteratureArticle Details
PMID: 7971991 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Three-dimensional structure of the adenine-specific DNA methyltransferase M.Taq I in complex with the cofactor S-adenosylmethionine.

Labahn J, Granzin J, Schluckebier G, Robinson DP, Jack WE, Schildkraut I, Saenger W

Abstract

The Thermus aquaticus DNA methyltransferase M.Taq I (EC 2.1.1.72) methylates N6 of adenine in the specific double-helical DNA sequence TCGA by transfer of --CH3 from the cofactor S-adenosyl-L-methionine. The x-ray crystal structure at 2.4-A resolution of this enzyme in complex with S-adenosylmethionine shows alpha/beta folding of the polypeptide into two domains of about equal size. They are arranged in the form of a C with a wide cleft suitable to accommodate the DNA substrate. The N-terminal domain is dominated by a nine-stranded beta-sheet; it contains the two conserved segments typical for N-methyltransferases which form a pocket for cofactor binding. The C-terminal domain is formed by four small beta-sheets and alpha-helices. The three-dimensional folding of M.Taq I is similar to that of the cytosine-specific Hha I methyltransferase, where the large beta-sheet in the N-terminal domain contains all conserved segments and the enzymatically functional parts, and the smaller C-terminal domain is less structured.

MeSH Terms
Bacterial Proteins/ultrastructure Base Sequence Binding Sites Crystallography, X-Ray DNA/chemistry Macromolecular Substances Molecular Sequence Data Protein Structure, Secondary Protein Structure, Tertiary Recombinant Proteins S-Adenosylmethionine/chemistry Site-Specific DNA-Methyltransferase (Adenine-Specific)/ultrastructure Thermus/enzymology
Chemicals
Bacterial Proteins Macromolecular Substances Recombinant Proteins S-Adenosylmethionine DNA Site-Specific DNA-Methyltransferase (Adenine-Specific)
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Labahn J
Institut für Kristallographie, Freie Universität, Berlin, Germany.
Granzin J
Schluckebier G
Robinson D P
Jack W E
Schildkraut I
Saenger W
References (20)
20 references, click to expand
  1. Kinetic and catalytic mechanism of HhaI methyltransferase.
    J Biol Chem. 1987 Apr 5;262(10):4778-86 PMID: 3558369
  2. Amino acid sequence arrangements of DNA-methyltransferases.
    Methods Enzymol. 1992;216:259-79 PMID: 1479903
  3. Sequence motifs characteristic of DNA[cytosine-N4]methyltransferases: similarity to adenine and cytosine-C5 DNA-methylases.
    Nucleic Acids Res. 1989 Dec 11;17(23):9823-32 PMID: 2690010
  4. Overexpression, purification and crystallization of BamHI endonuclease.
    Nucleic Acids Res. 1991 Apr 25;19(8):1825-9 PMID: 2030964
  5. Stereochemical studies of the C-methylation of deoxycytidine catalyzed by HhaI methylase and the N-methylation of deoxyadenosine catalyzed by EcoRI methylase.
    Arch Biochem Biophys. 1991 Feb 1;284(2):264-9 PMID: 1989510
  6. HhaI methyltransferase flips its target base out of the DNA helix.
    Cell. 1994 Jan 28;76(2):357-69 PMID: 8293469
  7. The sequence specificity domain of cytosine-C5 methylases.
    Nucleic Acids Res. 1991 Nov 25;19(22):6183-90 PMID: 1659688
  8. The corrected nucleotide sequences of the TaqI restriction and modification enzymes reveal a thirteen-codon overlap.
    Gene. 1992 Mar 1;112(1):91-5 PMID: 1551602
  9. Direct identification of the active-site nucleophile in a DNA (cytosine-5)-methyltransferase.
    Biochemistry. 1991 Nov 19;30(46):11018-25 PMID: 1932026
  10. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  11. Crystal structure of the HhaI DNA methyltransferase complexed with S-adenosyl-L-methionine.
    Cell. 1993 Jul 30;74(2):299-307 PMID: 8343957
  12. Predictive motifs derived from cytosine methyltransferases.
    Nucleic Acids Res. 1989 Apr 11;17(7):2421-35 PMID: 2717398
  13. How M.MspI and M.HpaII decide which base to methylate.
    Nucleic Acids Res. 1992 Sep 25;20(18):4811-6 PMID: 1408795
  14. PRISM: topologically constrained phased refinement for macromolecular crystallography.
    Acta Crystallogr D Biol Crystallogr. 1993 Sep 1;49(Pt 5):429-39 PMID: 15299502
  15. Cloning and sequence analysis of the genes coding for Eco57I type IV restriction-modification enzymes.
    Nucleic Acids Res. 1992 Nov 25;20(22):6051-6 PMID: 1334261
  16. DNA methylation: a phoenix rises.
    Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8761-2 PMID: 8415603
  17. On the mechanism of DNA-adenine methylase.
    J Biol Chem. 1988 Jun 5;263(16):7461-4 PMID: 3259576
  18. Crystallographic R factor refinement by molecular dynamics.
    Science. 1987 Jan 23;235(4787):458-60 PMID: 17810339
  19. Prediction of the occurrence of the ADP-binding beta alpha beta-fold in proteins, using an amino acid sequence fingerprint.
    J Mol Biol. 1986 Jan 5;187(1):101-7 PMID: 3959077
  20. Cloning, sequencing and expression of the Taq I restriction-modification system.
    Nucleic Acids Res. 1987 Dec 10;15(23):9781-96 PMID: 2827113
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1994-11-08
Pages
10957-61
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC45145
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]