Skip Navigation

This Article
Right arrow Print PDF (1808K)
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 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 (41)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Friedman, S.
Right arrow Articles by Ansari, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Friedman, S.
Right arrow Articles by Ansari, N.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1992, Vol. 20, No. 12 3241-3248
© 1992

MOLECULAR BIOLOGY

Binding of the EcoRII methyltransferase to 5-fluorocytosine-containing DNA. Isolation of a bound peptide

Stanley Friedman and Nasrin Ansari

Department of Pharmacology, SUNY Hearth Science Center at Brooklyn Box 29, 450 Clarkson Avenue, Brooklyn, NY 11203, USA

Received October 22, 1991. Revised May 12, 1992. Accepted May 12, 1992.

The properties of the interaction of 5-fluorocytosine-containing DNA with the EcoRII methyltransferase were studied. The DNA used was either a polymer synthesized in vitro, or a 20-mer containing one CCA/TGG sequence. The DNA could be methylated by the enzyme. In the process the enzyme formed a tight binding adduct with the DNA that could be identified by sodium dodecyl sulfate-polyacrylamide gel elctrophoresis. Enzyme activity was inhibited by this interaction. The 20-mer could be used to titrate the active site of the enzyme. The DNA polymer formed a tight binding complex that could be identified following digestion of the DNA with pancreatic deoxy-ribonuclease or micrococcal nuclease. A peptide-DNA adduct could be isolated after digestion of the EcoRII-DNA adduct with staphylococcal protease V8 by high pressure liquid chromatography and polyacryl-amide gel electrophoresis. Sequencing of the peptide indicated the DNA bound to a region of the protein that is conserved in all procaryotic DNA(cytosine-5)-methyltransferases. We have previously shown that this region contains a cysteine that can be photo-methylated with adenosylmethionine. This region, in addition to forming part of, or being adjacent to, the AdoMet binding site, also forms part of the DNA binding site.


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
 Proc. Natl. Acad. Sci. USAHome page
S. S. Smith, L. Niu, D. J. Baker, J. A. Wendel, S. E. Kane, and D. S. Joy
Nucleoprotein-based nanoscale assembly
PNAS, March 18, 1997; 94(6): 2162 - 2167.
[Abstract] [Full Text] [PDF]

Home page
 DevelopmentHome page
H Lei, S. Oh, M Okano, R Juttermann, K. Goss, R Jaenisch, and E Li
De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells
Development, January 10, 1996; 122(10): 3195 - 3205.
[Abstract] [PDF]

Home page
 Cold Spring Harb Symp Quant BiolHome page
X. Cheng, S. Kumar, S. Klimasauskas, and R.J. Roberts
Crystal Structure of the HhaI DNA Methyltransferase
Cold Spring Harb Symp Quant Biol, January 1, 1993; 58(0): 331 - 338.
[Abstract] [PDF]

Home page
 J. Biol. Chem.Home page
G. Vilkaitis, E. Merkiene, S. Serva, E. Weinhold, and S. Klimasauskas
The Mechanism of DNA Cytosine-5 Methylation. KINETIC AND MUTATIONAL DISSECTION OF HhaI METHYLTRANSFERASE
J. Biol. Chem., June 8, 2001; 276(24): 20924 - 20934.
[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.