Functional anatomy of polycomb and trithorax chromatin landscapes in Drosophila embryos

PLoS Biol. 2009 Jan 13;7(1):e13. doi: 10.1371/journal.pbio.1000013.

Abstract

Polycomb group (PcG) and trithorax group (trxG) proteins are conserved chromatin factors that regulate key developmental genes throughout development. In Drosophila, PcG and trxG factors bind to regulatory DNA elements called PcG and trxG response elements (PREs and TREs). Several DNA binding proteins have been suggested to recruit PcG proteins to PREs, but the DNA sequences necessary and sufficient to define PREs are largely unknown. Here, we used chromatin immunoprecipitation (ChIP) on chip assays to map the chromosomal distribution of Drosophila PcG proteins, the N- and C-terminal fragments of the Trithorax (TRX) protein and four candidate DNA-binding factors for PcG recruitment. In addition, we mapped histone modifications associated with PcG-dependent silencing and TRX-mediated activation. PcG proteins colocalize in large regions that may be defined as polycomb domains and colocalize with recruiters to form several hundreds of putative PREs. Strikingly, the majority of PcG recruiter binding sites are associated with H3K4me3 and not with PcG binding, suggesting that recruiter proteins have a dual function in activation as well as silencing. One major discriminant between activation and silencing is the strong binding of Pleiohomeotic (PHO) to silenced regions, whereas its homolog Pleiohomeotic-like (PHOL) binds preferentially to active promoters. In addition, the C-terminal fragment of TRX (TRX-C) showed high affinity to PcG binding sites, whereas the N-terminal fragment (TRX-N) bound mainly to active promoter regions trimethylated on H3K4. Our results indicate that DNA binding proteins serve as platforms to assist PcG and trxG binding. Furthermore, several DNA sequence features discriminate between PcG- and TRX-N-bound regions, indicating that underlying DNA sequence contains critical information to drive PREs and TREs towards silencing or activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / physiology*
  • Chromatin Immunoprecipitation
  • Chromosomal Proteins, Non-Histone / physiology*
  • Drosophila Proteins / physiology*
  • Drosophila melanogaster / embryology*
  • Gene Expression Regulation, Developmental / physiology
  • Polycomb Repressive Complex 1
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • Drosophila Proteins
  • Pc protein, Drosophila
  • trx protein, Drosophila
  • Polycomb Repressive Complex 1