title: Multiple distinct stimuli increase measured nucleosome occupancy around human promoters.
abstract: Nucleosomes can block access to transcription factors. Thus the precise localization of nucleosomes relative to transcription start sites and other factor binding sites is expected to be a critical component of transcriptional regulation. Recently developed microarray approaches have allowed the rapid mapping of nucleosome positions over hundreds of kilobases (kb) of human genomic DNA, although these approaches have not yet been widely used to measure chromatin changes associated with changes in transcription. Here, we use custom tiling microarrays to reveal changes in nucleosome positions and abundance that occur when hormone-bound glucocorticoid receptor (GR) binds to sites near target gene promoters in human osteosarcoma cells. The most striking change is an increase in measured nucleosome occupancy at sites spanning ∼1 kb upstream and downstream of transcription start sites, which occurs one hour after addition of hormone, but is lost at 4 hours. Unexpectedly, this increase was seen both on GR-regulated and GR-non-regulated genes. In addition, the human SWI/SNF chromatin remodeling factor (a GR co-activator) was found to be important for increased occupancy upon hormone treatment and also for low nucleosome occupancy without hormone. Most surprisingly, similar increases in nucleosome occupancy were also seen on both regulated and non-regulated promoters during differentiation of human myeloid leukemia cells and upon activation of human CD4+ T-cells. These results indicate that dramatic changes in chromatin structure over ∼2 kb of human promoters may occur genomewide and in response to a variety of stimuli, and suggest novel models for transcriptional regulation.
authors.affiliation: AstraZeneca R&D Boston, Waltham, Massachusetts, United States of America.
authors.forename: Chuong D
authors.initials: CD
authors.lastname: Pham
authors.forename: Hillel I
authors.initials: HI
authors.lastname: Sims
authors.forename: Trevor K
authors.initials: TK
authors.lastname: Archer
authors.forename: Gavin R
authors.initials: GR
authors.lastname: Schnitzler
mesh_terms: D001665:Binding Sites; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D018983:DNA Footprinting; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D015894:Genome, Human / Q000235:genetics; D005938:Glucocorticoids / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects / Q000235:genetics; D009707:Nucleosomes / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000187:drug effects; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D005938:Glucocorticoids; D009707:Nucleosomes; D011965:Receptors, Glucocorticoid; C445179:SWI-SNF-B chromatin-remodeling complex; D014157:Transcription Factors; D003907:Dexamethasone