Abstract
Specific DNA targets of chromatin-associated factors have been enzymatically marked in order to identify loci that are regulated at the level of chromatin organization in eukaryotes.
Highlights
Throughout differentiation, cells must activate some genes and silence others, and these states must be maintained during replication
Escherichia coli DNA adenine methyltransferase (Dam), which catalyzes methylation of adenine, was used as a reporter enzyme fused to Drosophila heterochromatin protein 1 (HP1)
HP1 became integrated into the chromatin structure and directed the Dam methyltransferase to HP1-associated DNA sequences
Summary
Throughout differentiation, cells must activate some genes and silence others, and these states must be maintained during replication. Existing techniques to identify protein-DNA interactions, such as chromatin immunoprecipitation and immunofluorescence, are technically difficult and are potentially subject to artifacts. It has proved extremely difficult to identify their targets in vitro This may be because they only recognize DNA that is already complexed with other proteins. One such factor, implicated in epigenetic phenomena in Drosophila, is the highly conserved heterochromatin protein 1 (HP1). It does not directly bind DNA but is predominantly associated with heterochromatic (highly condensed and repetitive) chromosomal regions, which are thought to have a silencing effect on gene expression. The authors describe a novel in vivo assay that has identified the DNA sequences that are (or have been) targeted by HP1 before, or at the time of, analysis
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