Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (209K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (11)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Kiss, A.
Right arrow Articles by Venetianer, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kiss, A.
Right arrow Articles by Venetianer, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 15 3188-3194
© 2001 Oxford University Press

Role of DNA minor groove interactions in substrate recognition by the M.SinI and M.EcoRII DNA (cytosine-5) methyltransferases

Antal Kiss*, György Pósfai, Gábor Zsurka, Tamás Raskó and Pál Venetianer

Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, PO Box 521, Szeged 6701, Hungary

The SinI and EcoRII DNA methyltransferases recognize sequences (GGA/TCC and CCA/TGG, respectively), which are characterized by an A/T ambiguity. Recognition of the A·T and T·A base pair was studied by in vitro methyltransferase assays using oligonucleotide substrates containing a hypoxanthine·C base pair in the central position of the recognition sequence. Both enzymes methylated the substituted oligonucleotide with an efficiency that was comparable to methylation of the canonical substrate. These observations indicate that M.SinI and M.EcoRII discriminate between their canonical recognition site and the site containing a G·C or a C·G base pair in the center of the recognition sequence (GGG/CCC and CCG/CGG, respectively) by interaction(s) in the DNA minor groove. M.SinI mutants displaying a decreased capacity to discriminate between the GGA/TCC and GGG/CCC sequences were isolated by random mutagenesis and selection for the relaxed specificity phenotype. These mutations led to amino acid substitutions outside the variable region, previously thought to be the sole determinant of sequence specificity. These observations indicate that A/T versus G/C discrimination is mediated by interactions between the large domain of the methyltransferase and the minor groove surface of the DNA.

* To whom correspondence should be addressed. Tel: +36 62 432 080; Fax: +36 62 433 506; Email: kissa{at}nucleus.szbk.u-szeged.hu Present address:Gábor Zsurka, Institute of Microbiology and Genetics, Vienna Biocenter, Dr Bohr-Gasse 9, A-1030 Vienna, Austria


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
M. Sokolowska, H. Czapinska, and M. Bochtler
Crystal structure of the {beta}{beta}{alpha}-Me type II restriction endonuclease Hpy99I with target DNA
Nucleic Acids Res., June 1, 2009; 37(11): 3799 - 3810.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
E. Timar, P. Venetianer, and A. Kiss
In Vivo DNA Protection by Relaxed-Specificity SinI DNA Methyltransferase Variants
J. Bacteriol., December 15, 2008; 190(24): 8003 - 8008.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
E. Timar, G. Groma, A. Kiss, and P. Venetianer
Changing the recognition specificity of a DNA-methyltransferase by in vitro evolution
Nucleic Acids Res., July 25, 2004; 32(13): 3898 - 3903.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
H. M. Cohen, D. S. Tawfik, and A. D. Griffiths
Altering the sequence specificity of HaeIII methyltransferase by directed evolution using in vitro compartmentalization
Protein Eng. Des. Sel., January 1, 2004; 17(1): 3 - 11.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y.-F. Lee, D. S. Tawfik, and A. D. Griffiths
Investigating the target recognition of DNA cytosine-5 methyltransferase HhaI by library selection using in vitro compartmentalisation
Nucleic Acids Res., November 15, 2002; 30(22): 4937 - 4944.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.