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Genome wide co-localization of Polycomb orthologs and their effects on gene expression in human fibroblasts

ABSTRACT: We have identfied antibodies that support chromatin immunoprecipitation (ChIP) of 11 components of Polycomb repressive complex 1 (PRC1) and shown that multiple variants of PRC1 associate with the same DNA. By performing ChIP-seq with antibodies against CBX6, CBX7, CBX8, RING1 and RING2, in two strains of human fibroblasts, we find that all five proteins co-localize at multiple sites in the genome. The binding profiles have inherent architecture that is conserved between the two strains, but there are also a substantial number of differences between the strains, possibly reflecting their anatomical position. The presence of PcG proteins does not equate with gene silencing and their genomic position does not change at senescence.

ORGANISM(S): Homo sapiens

PROVIDER: GSE40740 | GEO | 2013/12/19

SECONDARY ACCESSION(S): PRJNA174794

REPOSITORIES: GEO

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Genome wide co-localization of Polycomb orthologs and their effects on gene expression in human fibroblasts

Project description:We have identfied antibodies that support chromatin immunoprecipitation (ChIP) of 11 components of Polycomb repressive complex 1 (PRC1) and shown that multiple variants of PRC1 associate with the same DNA. By performing ChIP-seq with antibodies against CBX6, CBX7, CBX8, RING1 and RING2, in two strains of human fibroblasts, we find that all five proteins co-localize at multiple sites in the genome. The binding profiles have inherent architecture that is conserved between the two strains, but there are also a substantial number of differences between the strains, possibly reflecting their anatomical position. The presence of PcG proteins does not equate with gene silencing and their genomic position does not change at senescence. Chromatin immunoprecipitation and sequencing (ChIP-seq) in two strains of human fibroblasts using antibodies against three Pc orthologs (CBX6, CBX7 and CBX8) and both Psc orthologs (RING1 and 2). Validation by ChIP-PCR and re-ChIP. Strand specific RNA-seq and validation by qRT-PCR.

2013-12-19 | E-GEOD-40740 | biostudies-arrayexpress

Polycomb Cbx orthologs mediate the balance between Hematopoietic Stem Cell self-renewal and differentiation

2012-12-21 | GSE36658 | GEO

Polycomb Cbx orthologs mediate the balance between Hematopoietic Stem Cell self-renewal and differentiation

Project description:By genome-wide ChIP-seq analyses we show that Cbx7 and Cbx8 share ~95% of their targets in bone marrow cells. Cbx7 and Cbx8 have only ~200 targets that are not overlapping between the two proteins. These distinct Cbx7 and Cbx8 targets show reciprocal expression patterns along hematopoietic differentiation. Overexpression of empty vector (negative control), flag-Cbx7 and flag-Cbx8 in 5-FU treated bone marrow cells (progenitor and stem cells), followed by 1 week culture and subsequent ChIP using flag-M2 agarose beads.

2012-12-21 | E-GEOD-36658 | biostudies-arrayexpress

A Cbx8-containing Polycomb complex facilitates gene activation during ES cell differentiation [ChIP-Seq]

Project description:Polycomb proteins play an essential role in maintaining the repression of developmental genes in self-renewing embryonic stem cells. The exact mechanism allowing the derepression of polycomb target genes during cell differentiation remains unclear. Here, we show that several differentiation genes transiently recruit a Cbx8-containing Polycomb repressive complex (PRC) 1 during their early activation. Depletion of Cbx8 partially impairs the transcriptional activation of these genes. This correlates with a reduction in low but detectable levels of histone H3 lysine 27 acetylation. Prolonged gene activation results in eviction of PRC1 despite persisting H3K27me3. The composition of PRC1 is highly modular and changes when ES cells commit to differentiation. We further demonstrate that the exchange of Cbx7 for Cbx8 is required for the effective activation of differentiation genes. Taken together our results establish a function for a Cbx8-containing complex in facilitating the transition from a Polycomb-repressed chromatin state to an active state. As this affects several key regulatory differentiation genes this mechanism is likely to contribute to the robust execution of differentiation programs. Examination of cbx8 in ES E14 mouse cells in 2 condition before and after 72h stimulation with retinoic acid compared with IgG

2014-11-04 | E-GEOD-54052 | biostudies-arrayexpress

A Cbx8-containing Polycomb complex facilitates gene activation during ES cell differentiation [ChIP-Seq]

2014-11-04 | GSE54052 | GEO

A Cbx8-containing Polycomb complex facilitates gene activation during ES cell differentiation [Agilent Array]

2014-11-04 | GSE54051 | GEO

RING1 links retinoic acid signaling to the early proximal-distal specification of forelimb bud via Meis2 repression (ChIP-Seq)

Project description:Polycomb group (PcG) proteins play a pivotal role in silencing of development-related genes and contribute to maintain various stem and precursor cells and regulate their differentiation. However, it is not well understood how PcG factors regulate dynamic and complex morphogenetic processes particularly in mammals. In this study, we focused on proximal-distal (PD) patterning of forelimb bud to elucidate how PcG factors contribute to regulation of morphogenetic processes that depends on developmental signals. Depletion of RING1 proteins, which are common components of both canonical and variant Polycomb repressive complex-1 (PRC1), led to dramatic deficiencies in forelimb formation. By using Ring1-deficient forelimb buds, we revealed the early defects in distal specification, which is due to the functional coupling of RING1 and retinoic acid (RA) signaling. RING1 activity is shown to antagonize with RA signals at Meis2 and likely Meis1, which are superior suppressors of distal formation program, in the prospective distal region of outgrowing forelimb bud. This study exhibits a first example showing how PcG factors interplay with developmental signals to mediate compartmentalization of elongating anlagen by regulating the expression of developmental genes.

2015-12-28 | GSE70076 | GEO

RING1 links retinoic acid signaling to the early proximal-distal specification of forelimb bud via Meis2 repression (ChIP-chip)

2015-12-28 | GSE70074 | GEO

RING1 links retinoic acid signaling to the early proximal-distal specification of forelimb bud via Meis2 repression (mRNA)

2015-12-28 | GSE70075 | GEO

A Cbx8-containing Polycomb complex facilitates gene activation during ES cell differentiation [Agilent Array]

Project description:Polycomb proteins play an essential role in maintaining the repression of developmental genes in self-renewing embryonic stem cells. The exact mechanism allowing the derepression of polycomb target genes during cell differentiation remains unclear. Here, we show that several differentiation genes transiently recruit a Cbx8-containing Polycomb repressive complex (PRC) 1 during their early activation. Depletion of Cbx8 partially impairs the transcriptional activation of these genes. This correlates with a reduction in low but detectable levels of histone H3 lysine 27 acetylation. Prolonged gene activation results in eviction of PRC1 despite persisting H3K27me3. The composition of PRC1 is highly modular and changes when ES cells commit to differentiation. We further demonstrate that the exchange of Cbx7 for Cbx8 is required for the effective activation of differentiation genes. Taken together our results establish a function for a Cbx8-containing complex in facilitating the transition from a Polycomb-repressed chromatin state to an active state. As this affects several key regulatory differentiation genes this mechanism is likely to contribute to the robust execution of differentiation programs. Four biological replicates were used for each condition (untreated and after 72h of retinoic acid stimulation) and samples were prepared and hybridized to SurePrint G3 Gene Expression Microarrays (Agilent technologies) following the supplier's instructions. Analyses were essentially performed as described (Uribesalgo et al. 2011) selecting differentially expressed probes with a FDR of 0.05 and fold change of > 1.5.

2014-11-04 | E-GEOD-54051 | biostudies-arrayexpress

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