Silencing of human polycomb target genes is associated with methylation of histone H3 Lys 27

Kirmizis, Antonis; Bartley, Stephanie M.; Kuzmichev, Andrei; Margueron, Raphaël; Reinberg, Danny; Green, Roland; Farnham, Peggy J.

doi:10.1101/gad.1200204

Cited by 453 publications

(410 citation statements)

References 38 publications

Supporting

Mentioning

394

Contrasting

Unclassified

Order By: Relevance

“…MYT1 (myelin-transcription-factor 1), a well-established Polycomb target gene (Kirmizis et al, 2004), served as a positive control and glyceraldehyde-3 phosphate dehydrogenase (GAPDH) as a negative control. Importantly, whereas the MSMB gene was heavily trimethylated on H3K27 in PC-3 cells, which hardly express PSP94, this gene was only mildly trimethylated on H3K27 in LNCaP cells, which express a lot of PSP94 (Figure 1a and c).…”

Section: Introduction Results and Discussionmentioning

confidence: 99%

The gene encoding the prostatic tumor suppressor PSP94 is a target for repression by the Polycomb group protein EZH2

et al. 2007

View full text Add to dashboard Cite

BelgiumPSP94, for prostatic secretory protein of 94 amino acids, is secreted by the prostate gland and functions as a suppressor of tumor growth and metastasis. The expression of PSP94 is lost in advanced, hormone-refractory prostate cancer and this correlates with an increased expression of the Polycomb protein EZH2 (enhancer of zeste homolog 2), which represses transcription via trimethylation of histone H3 on Lys27 (H3K27). We show here that these events are causally related and that the MSMB gene, which encodes PSP94, is trimethylated on H3K27 in androgen-refractory, but not in androgensensitive prostate cancer cells. Chromatin immunoprecipitation experiments confirmed an association of EZH2 with the MSMB gene. The RNAi-mediated knockdown of EZH2 resulted in a loss of H3K27 trimethylation and an increased expression of the MSMB gene. Conversely, the overexpression of EZH2 was associated with a decreased expression of the MSMB gene. We also demonstrate that MSMB is additionally repressed in androgen-refractory prostate cancer cells by the hypoacetylation of histone H3K9 and the hypermethylation of a CpG island in the promoter region. Our data disclose a hitherto unexplored link between the putative oncogene EZH2 and the tumor suppressor PSP94, and show that MSMB is silenced by EZH2 in advanced prostate cancer cells.

show abstract

Section: Introduction Results and Discussionmentioning

confidence: 99%

The gene encoding the prostatic tumor suppressor PSP94 is a target for repression by the Polycomb group protein EZH2

et al. 2007

View full text Add to dashboard Cite

show abstract

“…ChIP/chip analysis was carried out as described by others [11][12][13][14]. In brief, DNA was isolated by ChIP methodology followed by ligation mediated PCR as described by Oberley et al [14].…”

Section: Tiled Oligonucleotide Microarray Analysismentioning

confidence: 99%

Multiple enhancer regions located at significant distances upstream of the transcriptional start site mediate RANKL gene expression in response to 1,25-dihydroxyvitamin D3

Kim

Yamazaki

Zella

et al. 2007

The Journal of Steroid Biochemistry and Molecular Biology

View full text Add to dashboard Cite

One of the primary regulators of Receptor activator of NF-κB ligand (RANKL) is 1,25-dihydoxyvitamin D 3 (1,25(OH) 2 D 3 ). To elucidate the mechanism whereby 1,25(OH) 2 D 3 activates RANKL expression we screened some 300 kb of the RANKL gene locus using a ChIP on chip analysis and identified five potential regulatory regions lying significant distances upstream of the transcription start site (TSS), the farthest over 70 kb from the TSS. A direct ChIP analysis confirmed the presence of the VDR/RXR heterodimer at these sites. The binding of the VDR was associated with histone modification and enhanced entry of RNA polymerase II, indicating an important functional consequence to the localization of these transcription factors in response to 1,25 (OH) 2 D 3 . The region -76kb upstream from the TSS, termed D5, was capable of mediating VDRdependent transcriptional output in response to 1,25(OH) 2 D 3 in luciferase assays. The identified VDRE in this region was able to confer dramatic 1,25(OH) 2 D 3 sensitivity to heterologous promoters. This region was highly evolutionarily conserved and functionally active in the human RANKL gene as well. We propose that the RANKL gene is regulated via multiple enhancers that while located at significant distances from the TSS, likely form a chromatin hub centered on the RankL promoter.

show abstract

“…CGIs are evolutionarily conserved elements corresponding to the promoters and regulatory regions of more than 50% of the genes in the human genome (6). CGI arrays have been used in a number of studies to identify regulatory elements bound directly by transcription factors, including E2F, MYC, and Suz12 (40,53,60,82). To date, there have been several publications describing global ChIP-based approaches for identifying new p53-regulated genes, but none have used CGI arrays to do so (9,27,36,81).…”

mentioning

confidence: 99%

Functional Analysis of p53 Binding under Differential Stresses

Krieg

Hammond

Giaccia

2006

Molecular and Cellular Biology

View full text Add to dashboard Cite

Hypoxia and DNA damage stabilize the p53 protein, but the subsequent effect that each stress has on transcriptional regulation of known p53 target genes is variable. We have used chromatin immunoprecipitation followed by CpG island (CGI) microarray hybridization to identify promoters bound by p53 under both DNA-damaging and non-DNA-damaging conditions in HCT116 cells. Using gene-specific PCR analysis, we have verified an association with CGIs of the highest enrichment (>2.5-fold) (REV3L, XPMC2H, HNRPUL1, TOR1AIP1, glutathione peroxidase 1, and SCFD2), with CGIs of intermediate enrichment (>2.2-fold) (COX7A2L, SYVN1, and JAG2), and with CGIs of low enrichment (>2.0-fold) (MYC and PCNA). We found little difference in promoter binding when p53 is stabilized by these two distinctly different stresses. However, expression of these genes varies a great deal: while a few genes exhibit classical induction with adriamycin, the majority of the genes are unchanged or are mildly repressed by either hypoxia or adriamycin. Further analysis using p53 mutated in the core DNA binding domain revealed that the interaction of p53 with CGIs may be occurring through both sequence-dependent and -independent mechanisms. Taken together, these experiments describe the identification of novel p53 target genes and the subsequent discovery of distinctly different expression phenomena for p53 target genes under different stress scenarios.The tumor microenvironment is a major factor influencing tumor growth, metastatic potential, and response to chemotherapeutic drugs and radiation therapy (7). The hypoxia-inducible factor (HIF) family of transcription factors regulates the expression of key enzymes in anaerobic glycolysis and proangiogenic factors (i.e., vascular endothelial growth factor), aiding cells in adapting to a low-oxygen environment (15). Thus, a tumor adapts to hypoxia first by sustaining itself via anaerobic metabolism and then by stimulating angiogenesis to acquire more nutrients and oxygen. However, the resulting vasculature tends to have aberrant structures with irregular blood flow and leaky walls, paradoxically creating regions of transient hypoxia and reoxygenation (76). Reoxygenation after hypoxia causes DNA damage, exacerbating genomic instability (25). The porous structure of the tumor vasculature may also provide an escape route for metastasizing cells. Repeated cycles of hypoxia, vascular formation, and reoxygenation select for cells that are more likely to survive in a restrictive environment. The consequences of this process are cancer cells that are more aggressive, more likely to metastasize, and more likely to be resistant to radiation and chemotherapy. Intimately tied to this process is the selection for cells that have lost the expression of functional p53 (19).A variety of cellular stresses, including DNA damage, oncogenic transformation, and hypoxia, stabilize the p53 protein by activating a host of stress-inducible kinases that phosphorylate p53 and MDM2, resulting in increased levels of p53 (17). Stabilized ...

show abstract

Silencing of human polycomb target genes is associated with methylation of histone H3 Lys 27

Cited by 453 publications

References 38 publications

The gene encoding the prostatic tumor suppressor PSP94 is a target for repression by the Polycomb group protein EZH2

The gene encoding the prostatic tumor suppressor PSP94 is a target for repression by the Polycomb group protein EZH2

Multiple enhancer regions located at significant distances upstream of the transcriptional start site mediate RANKL gene expression in response to 1,25-dihydroxyvitamin D3

Functional Analysis of p53 Binding under Differential Stresses

Contact Info

Product

Resources

About