Chromatin profiling using targeted DNA adenine methyltransferase.
Chromatin is the highly complex structure consisting of DNA and hundreds of associated proteins. Most chromatin proteins exert their regulatory and structural functions by binding to specific chromosomal loci. Knowledge of the identity of these in vivo target loci is essential for the understanding of the functions and mechanisms of action of chromatin proteins. We report here large-scale mapping of in vivo binding sites of chromatin proteins, using a novel approach based on a combination of targeted DNA methylation and microarray technology. We show that three distinct chromatin proteins in Drosophila melanogaster cells each associate with specific sets of genes. HP1 binds predominantly to pericentric genes and transposable elements. GAGA factor associates with euchromatic genes that are enriched in (GA)n motifs. A Drosophila homolog of Saccharomyces cerevisiae Sir2p is associated with several active genes and is excluded from heterochromatin. High-resolution, genome-wide maps of target loci of chromatin proteins ('chromatin profiles') provide new insights into chromatin structure and gene regulation.
van Steensel
B
B
Howard Hughes Medical Institute, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA. steesel@chem.uva.nl
Delrow
J
J
Henikoff
S
S
D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D019175:DNA Methylation; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)
D002843:Chromatin; C058693:Dam methyltransferase; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)