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Multiple roles of H2A.Z in regulating promoter chromatin architecture in human cells (Mnase-seq & ChIP-seq)

ABSTRACT: We employed an MNase-Transcription Start Site Sequence Capture method to map and determine the accessibility of all nucleosomes, including those that contain H2A.Z, at high coverage for all human Pol II promoters. We uncovered unexpected features of nucleosomal organization in epithelial cells and following transition to a mesenchymal and malignant state. In contrast to the prevailing model, we observe many different types of active and inactive promoter structures that differ in their nucleosome organization and sensitivity to MNase digestion. Further, we found that H2A.Z has an important role in the assembly of repressed promoters into an inaccessible state. Finally, we uncovered a new promoter type in which a stable H2A.Z nucleosome occupies the TSS of an active promoter.

ORGANISM(S): Homo sapiens

PROVIDER: GSE134297 | GEO | 2020/06/01

REPOSITORIES: GEO

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Multiple roles of H2A.Z in regulating promoter chromatin architecture in human cells (RNA-seq)

Project description:We employed an MNase-Transcription Start Site Sequence Capture method to map and determine the accessibility of all nucleosomes, including those that contain H2A.Z, at high coverage for all human Pol II promoters. We uncovered unexpected features of nucleosomal organization in epithelial cells and following transition to a mesenchymal and malignant state. In contrast to the prevailing model, we observe many different types of active and inactive promoter structures that differ in their nucleosome organization and sensitivity to MNase digestion. Further, we found that H2A.Z has an important role in the assembly of repressed promoters into an inaccessible state. Finally, we uncovered a new promoter type in which a stable H2A.Z nucleosome occupies the TSS of an active promoter.

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Canonical nucleosome organization at promoters forms during genome activation

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Project description:The position of nucleosomes influences DNA accessibility to DNA-binding proteins. Genome-wide nucleosome profiles often report the observation of a canonical nucleosome organization at gene promoters where arrays of well-positioned nucleosomes emanate from nucleosome-depleted regions. It is unclear how this canonical promoter nucleosome organization forms and how it is related to transcription activation and the establishment of histone marks during development. Here we report the genome-wide organization of nucleosomes during zebrafish embryogenesis and show that well-positioned nucleosome arrays appear in thousands of promoters during the activation of the zygotic genome. The formation of canonical promoter nucleosome organization cannot be explained by DNA sequence preference, and is independent of transcription and the presence of RNA polymerase II, but strongly correlates with the presence of Histone H3 Lysine 4 trimethylation (H3K4me3). Our study further suggests that promoter nucleosome structure primes genes to future transcription activation. To determine whether the occlusions are consistent in mammalian pluripotent cells, we performed the same analyses in mouse embryonic stem cells and found similar relationships. MNase-seq to generate nucleosome organization in mouse embryonic stem cell (J1)

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Effects of MBD3 depletion on nucleosome occupancy in MCF-7 cells

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