Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/31406
Share/Export:
logo share SHARE BASE
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE
Title

Dam methylase from Escherichia coli: kinetic studies using modified DNA oligomers: hemimethylated substrates

AuthorsMarzabal, Stephane; DuBois, Stephen; Thielking, Vera; Cano, Ángeles; Eritja Casadellà, Ramón CSIC ORCID ; Guschlbauer, Wilhem
Issue Date25-Sep-1995
PublisherOxford University Press
CitationNucleic Acids Research 23(18): 3648-3655 (1995)
AbstractWe have measured steady-state kinetics of the N6-adenine methyltransferase Dam Mtase using as substrates non-selfcomplementary tetradecamer duplexs (d[GCCGGATCTAGACG]-d[CGTCTAGATCC-GGC]) containing the hemimethylated GATC target sequence in one or the other strand and modifications in the GATC target sequence of the complementary strands. Modifications included substitution of guanine by hypoxanthine (I), thymine by uracil (U) or 5-ethyl-uracil (E) and adenine by 2,6-diamino-purine (D). Thermodynamic parameters were obtained from the concentration dependence of the melting temperature (Tm) of the duplexes. Large differences in DNA methylation of duplexes containing single dI for dG substitution of the Dam recognition site were observed compared with the canonical substrate, if the substitution involved the top strand (on the G.C rich side). Substitution in either strand by uracil (dU) or 5-ethyluracil (dE) resulted in small perturbation of the methylation patterns. When 2,6-diamino-purine (dD) replaced the adenine to be methylated, small, but significant methylation was observed. The kinetic parameters of the methylation reaction were compared with the thermodynamic free energies and significant correlation was observed.
Description8 pages, 6 figuras, 2 tables.-- PMID: 7478992 [PubMed].-- PMCID: PMC307261.
Publisher version (URL)http://www.ncbi.nlm.nih.gov/pmc/articles/PMC307261
URIhttp://hdl.handle.net/10261/31406
ISSN0305-1048
E-ISSN1362-4962
Appears in Collections:(IDAEA) Artículos

Files in This Item:
File Description SizeFormat
Marzabal_Stephane_et_al.pdf1,46 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work

Page view(s)

272
checked on May 22, 2022

Download(s)

98
checked on May 22, 2022

Google ScholarTM

Check


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.