Tiling histone H3 lysine 4 and 27 methylation in zebrafish using high-density microarrays

PLoS One. 2010 Dec 20;5(12):e15651. doi: 10.1371/journal.pone.0015651.

Abstract

Background: Uncovering epigenetic states by chromatin immunoprecipitation and microarray hybridization (ChIP-chip) has significantly contributed to the understanding of gene regulation at the genome-scale level. Many studies have been carried out in mice and humans; however limited high-resolution information exists to date for non-mammalian vertebrate species.

Principal findings: We report a 2.1-million feature high-resolution Nimblegen tiling microarray for ChIP-chip interrogations of epigenetic states in zebrafish (Danio rerio). The array covers 251 megabases of the genome at 92 base-pair resolution. It includes ∼15 kb of upstream regulatory sequences encompassing all RefSeq promoters, and over 5 kb in the 5' end of coding regions. We identify with high reproducibility, in a fibroblast cell line, promoters enriched in H3K4me3, H3K27me3 or co-enriched in both modifications. ChIP-qPCR and sequential ChIP experiments validate the ChIP-chip data and support the co-enrichment of trimethylated H3K4 and H3K27 on a subset of genes. H3K4me3- and/or H3K27me3-enriched genes are associated with distinct transcriptional status and are linked to distinct functional categories.

Conclusions: We have designed and validated for the scientific community a comprehensive high-resolution tiling microarray for investigations of epigenetic states in zebrafish, a widely used developmental and disease model organism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / chemistry
  • DNA Methylation
  • Epigenesis, Genetic
  • Fibroblasts / cytology
  • Gene Expression Regulation
  • Histones / chemistry*
  • Histones / genetics
  • Humans
  • Immunoprecipitation
  • Lysine / chemistry*
  • Methylation
  • Mice
  • Oligonucleotide Array Sequence Analysis / methods*
  • Promoter Regions, Genetic
  • Zebrafish

Substances

  • Chromatin
  • Histones
  • Lysine