Polycomb group repression reduces DNA accessibility

The Polycomb group proteins are responsible for long-term repression of a n umber of genes in Drosophila melanogaster, including the homeotic genes of the bithorax complex. The Polycomb protein is thought to alter the chromati n structure of its target genes, but there has been little direct evidence for this model. In this study, the chromatin structure of the bithorax comp lex was probed with three separate assays for DNA accessibility: (i) activa tion of polymerase II (Pot II) transcription by Gal4, (ii) transcription by the bacteriophage T7 RNA polymerase (T7RNAP), and (iii) FLP-mediated site- specific recombination. All three processes are restricted or blocked in Po lycomb-repressed segments. In contrast, control test sites outside of the b ithorax complex permitted Gal4, T7RNAP, and FLP activities throughout the e mbryo. Several P insertions in the bithorax complex were tested, providing evidence that the Polycomb-induced effect is widespread over target genes. This accessibility effect is similar to that seen for SIR silencing in Sacc haromyces cerevisiae. In contrast to SIR silencing, however, episomes excis ed from Polycomb-repressed chromosomal sites do not show an altered superhe lix density.