title: Activation of Bmp2-Smad1 signal and its regulation by coordinated alteration of H3K27 trimethylation in Ras-induced senescence.
abstract: Cellular senescence involves epigenetic alteration, e.g. loss of H3K27me3 in Ink4a-Arf locus. Using mouse embryonic fibroblast (MEF), we here analyzed transcription and epigenetic alteration during Ras-induced senescence on genome-wide scale by chromatin immunoprecipitation (ChIP)-sequencing and microarray. Bmp2 was the most activated secreted factor with H3K4me3 gain and H3K27me3 loss, whereas H3K4me3 loss and de novo formation of H3K27me3 occurred inversely in repression of nine genes, including two BMP-SMAD inhibitors Smad6 and Noggin. DNA methylation alteration unlikely occurred. Ras-activated cells senesced with nuclear accumulation of phosphorylated SMAD1/5/8. Senescence was bypassed in Ras-activated cells when Bmp2/Smad1 signal was blocked by Bmp2 knockdown, Smad6 induction, or Noggin induction. Senescence was induced when recombinant BMP2 protein was added to Bmp2-knocked-down Ras-activated cells. Downstream Bmp2-Smad1 target genes were then analyzed genome-wide by ChIP-sequencing using anti-Smad1 antibody in MEF that was exposed to BMP2. Smad1 target sites were enriched nearby transcription start sites of genes, which significantly correlated to upregulation by BMP2 stimulation. While Smad6 was one of Smad1 target genes to be upregulated by BMP2 exposure, Smad6 repression in Ras-activated cells with increased enrichment of Ezh2 and gain of H3K27me3 suggested epigenetic disruption of negative feedback by Polycomb. Among Smad1 target genes that were upregulated in Ras-activated cells without increased repressive mark, Parvb was found to contribute to growth inhibition as Parvb knockdown lead to escape from senescence. It was revealed through genome-wide analyses in this study that Bmp2-Smad1 signal and its regulation by harmonized epigenomic alteration play an important role in Ras-induced senescence.
authors.affiliation: Genome Science Division, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. kaneda@genome.rcast.u-tokyo.ac.jp
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kaneda
authors.forename: Takanori
authors.initials: T
authors.lastname: Fujita
authors.forename: Motonobu
authors.initials: M
authors.lastname: Anai
authors.forename: Shogo
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Genta
authors.initials: G
authors.lastname: Nagae
authors.forename: Masato
authors.initials: M
authors.lastname: Morikawa
authors.forename: Shingo
authors.initials: S
authors.lastname: Tsuji
authors.forename: Masanobu
authors.initials: M
authors.lastname: Oshima
authors.forename: Kohei
authors.initials: K
authors.lastname: Miyazono
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aburatani
mesh_terms: D000185:Actinin / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D016922:Cellular Senescence / Q000235:genetics*; D019175:DNA Methylation; D000071221:Enhancer of Zeste Homolog 2 Protein; D044127:Epigenesis, Genetic; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D011905:Genes, ras / Q000235:genetics; D016678:Genome; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D063151:Polycomb Repressive Complex 2; D015398:Signal Transduction; D051898:Smad1 Protein / Q000378:metabolism*; D051905:Smad6 Protein / Q000235:genetics / Q000378:metabolism; D000099176:Noggin Protein
chemical_list: D000185:Actinin; D055396:Bone Morphogenetic Protein 2; D002352:Carrier Proteins; D000071221:Enhancer of Zeste Homolog 2 Protein; D011495:Histone-Lysine N-Methyltransferase; D063151:Polycomb Repressive Complex 2; D051898:Smad1 Protein; D051905:Smad6 Protein; D000099176:Noggin Protein; C525719:Bmp2 protein, mouse; C500559:Parvb protein, mouse; C497401:Smad1 protein, mouse; C497803:Smad6 protein, mouse; C498410:Ezh2 protein, mouse
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