Identification of Protein-DNA Contacts with Dimethyl Sulfate

Methylation Protection and Methylation Interference
  • Peter E. Shaw
  • A. Francis Stewart
Part of the Methods in Molecular Biology™ book series (MIMB, volume 30)

Abstract

Dimethyl sulfate (DMS) is an effective and widely used probe for sequence-specific protein-DNA interactions. It is the only probe routinely used both for in vitro (methylation protection, methylation interference) and in vivo (DMS genomic footprinting) applications since it rapidly reacts with DNA at room temperature and readily penetrates intact cells (1). DMS predominantly methylates the 7-nitrogen of guanine and 3-nitrogen of adenine. Thus, reactivity with G residues occurs in the major groove and with A residues in the minor groove. In standard Maxam and Gilbert protocols (2), the methylated bases are subsequently converted to strand breaks and displayed on sequencing gels.

Keywords

Vortex EDTA Electrophoresis Isopropanol Adenine 

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Copyright information

© Humana Press Inc. 1994

Authors and Affiliations

  • Peter E. Shaw
    • 1
  • A. Francis Stewart
    • 2
  1. 1.Max Planck Institüt für ImmunbiologieFreiburg-ZähringenGermany
  2. 2.European Molecular Biology LaboratoriesHeidelbergGermany

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