Mapping in vivo protein-DNA interactions in plants by DamID, a DNA adenine methylation-based method

Methods Mol Biol. 2011:754:307-21. doi: 10.1007/978-1-61779-154-3_18.

Abstract

DamID (DNA adenine methylation identification) is an adenine methylation-based tagging method designed to map protein-DNA interactions in vivo. DamID, an alternative method to chromatin immunoprecipitation (ChIP), is based on the covalent linking of a "fingerprint" in the vicinity of the DNA-binding sites of the protein of interest. The fingerprints can be further mapped by simple molecular approaches. First developed by van Steensel's group in Drosophila melanogaster, DamID was successfully adapted to Arabidopsis thaliana, and its feasibility demonstrated by using the well-known yeast GAL4 transcription factor. The method was further used to establish a genome-wide map of the target sites of LHP1, a regulatory chromatin protein in A. thaliana.

MeSH terms

  • Adenine / metabolism
  • Arabidopsis / genetics
  • Binding Sites / genetics
  • Chromatin
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • DNA Methylation*
  • DNA, Plant / chemistry
  • DNA, Plant / genetics
  • DNA, Plant / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Genome, Plant
  • Polymerase Chain Reaction
  • Protein Interaction Mapping / methods*
  • Sequence Tagged Sites
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • DNA, Plant
  • DNA-Binding Proteins
  • Transcription Factors
  • like heterochromatin protein 1
  • Adenine