title: The CHD3 remodeler PICKLE associates with genes enriched for trimethylation of histone H3 lysine 27.
abstract: In Arabidopsis (Arabidopsis thaliana), the ATP-dependent chromatin remodeler PICKLE (PKL) determines expression of genes associated with developmental identity. PKL promotes the epigenetic mark trimethylation of histone H3 lysine 27 (H3K27me3) that facilitates repression of tissue-specific genes in plants. It has previously been proposed that PKL acts indirectly to promote H3K27me3 by promoting expression of the POLYCOMB REPRESSIVE COMPLEX2 complex that generates H3K27me3. We undertook expression and chromatin immunoprecipitation analyses to further characterize the contribution of PKL to gene expression and developmental identity. Our expression data support a critical and specific role for PKL in expression of H3K27me3-enriched loci but do not support a role for PKL in expression of POLYCOMB REPRESSIVE COMPLEX2. Moreover, our chromatin immunoprecipitation data reveal that PKL protein is present at the promoter region of multiple H3K27me3-enriched loci, indicating that PKL directly acts on these loci. In particular, we find that PKL is present at LEAFY COTYLEDON1 and LEAFY COTYLEDON2 during germination, which is when PKL acts to repress these master regulators of embryonic identity. Surprisingly, we also find that PKL is present at the promoters of actively transcribed genes that are ubiquitously expressed such as ACTIN7 and POLYUBIQUITIN10 that do not exhibit PKL-dependent expression. Taken together, our data contravene the previous model of PKL action and instead support a direct role for PKL in determining levels of H3K27me3 at repressed loci. Our data also raise the possibility that PKL facilitates a common chromatin remodeling process that is not restricted to H3K27me3-enriched regions.
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: Heng
authors.initials: H
authors.lastname: Zhang
authors.forename: Brett
authors.initials: B
authors.lastname: Bishop
authors.forename: Whitney
authors.initials: W
authors.lastname: Ringenberg
authors.forename: William M
authors.initials: WM
authors.lastname: Muir
authors.forename: Joe
authors.initials: J
authors.lastname: Ogas
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D004265:DNA Helicases; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D056426:Genetic Loci; D018525:Germination; D006657:Histones / Q000378:metabolism*; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D008745:Methylation; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D063151:Polycomb Repressive Complex 2; D025821:Polyubiquitin / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D029681:Arabidopsis Proteins; D006657:Histones; C574170:PRC2 protein, Arabidopsis; D012097:Repressor Proteins; D025821:Polyubiquitin; D063151:Polycomb Repressive Complex 2; C402442:AT2G25170 protein, Arabidopsis; D004265:DNA Helicases; D008239:Lysine