DNA binding discrimination of the murine DNA cytosine-C5 methyltransferase

Journal of Molecular Biology
J FlynnN Reich

Abstract

Mammalian DNA cytosine-C5 methyltransferase modifies the CpG dinucleotide in the context of many different genomic sequences. A rigorous DNA binding assay was developed for the murine enzyme and used to define how sequences flanking the CpG dinucleotide affect the stability of the enzyme:DNA complex. Oligonucleotides containing a single CpG site form reversible 1:1 complexes with the enzyme that are sequence-specific. A guanine/cytosine-rich 30 base-pair sequence, a mimic of the GC-box cis-element, bound threefold more tightly than an adenine/thymine-rich sequence, a mimic of the cyclic AMP responsive element. However, the binding discrimination between hemi- and unmethylated forms of these DNA substrates was small, as we previously observed at the K(m)DNA level (Biochemistry, 35, 7308-7315 (1996)). Single-stranded substrates are bound much more weakly than double-stranded DNA forms. An in vitro screening method was used to select for CpG flanking sequence preferences of the DNA methyltransferase from a large, divergent population of DNA substrates. After five iterative rounds of increasing selective pressure, guanosine/cytosine-rich sequences were abundant and contributed to binding stabilization for at least 12 base-pairs on ...Continue Reading

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Citations

Feb 28, 2006·Nucleic Acids Research·Rachna GoyalAlbert Jeltsch
May 21, 2009·Biological Chemistry·Ernst G MalyginStanley Hattman
Jun 25, 2009·Molekuliarnaia biologiia·A A EvdokimovE G Malygin
Jan 5, 2014·Genetics and Molecular Biology·Thiago da Silva PaivaInácio Domingos da Silva-Neto
Dec 13, 2002·The Journal of Biological Chemistry·James FlynnNorbert O Reich
Apr 4, 2018·Nature Genetics·Wanding ZhouBenjamin P Berman
Jul 6, 2005·Biochemistry·Zeljko M Svedruzić, Norbert O Reich
Nov 9, 2005·Biochemistry·Zeljko M Svedruzić, Norbert O Reich
Sep 27, 2003·Biochemical and Biophysical Research Communications·Brandon E Aubol, Norbert O Reich
Jan 13, 2004·Biochemical and Biophysical Research Communications·Srivani SistlaDesirazu N Rao

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