Tgfβ signaling directly induces Arf promoter remodeling by a mechanism involving Smads 2/3 and p38 MAPK

Yanbin Zheng, Yi D. Zhao, Melissa Gibbons, Tatiana Abramova, Patricia Y. Chu, John D. Ash, John M. Cunningham, Stephen X. Skapek

Research output: Contribution to journalArticle

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Abstract

We have investigated how the Arf gene product, p19Arf, is activated by Tgfβ during mouse embryo development to better understand how this important tumor suppressor is controlled. Taking advantage of new mouse models, we provide genetic evidence that Arf lies downstream of Tgfβ signaling in cells arising from the Wnt1-expressing neural crest and that the anti-proliferative effects of Tgfβ depend on Arf in vivo. Tgfβ1, -2, and -3 (but not BMP-2, another member of the Tgfβ superfamily) induce p19Arf expression in wild type mouse embryo fibroblasts (MEFs), and they enhance Arf promoter activity in ArflacZ/lacZ MEFs. Application of chemical inhibitors of Smad-dependent and -independent pathways show that SB431542, a Tgfβ type I receptor (TβrI) inhibitor, and SB203580, a p38 MAPK inhibitor, impede Tgfβ2 induction of Arf. Genetic studies confirm the findings; transient knockdown of Smad2, Smad3, or p38 MAPK blunt Tgfβ2 effects, as does Cre recombinase treatment of Tgfbr2fl/fl MEFs to delete Tgfβ receptor II. Chromatin immunoprecipitation reveals that Tgfβ rapidly induces Smads 2/3 binding and histone H3 acetylation at genomic DNA proximal to Arf exon 1β. This is followed by increased RNA polymerase II binding and progressively increased Arf primary and mature transcripts from 24 through 72 h, indicating that increased transcription contributes to p19Arf increase. Last, Arf induction by oncogenic Ras depends on p38 MAPK but is independent of TβrI activation of Smad 2. These findings add to our understanding of how developmental and tumorigenic signals control Arf expression in vivo and in cultured MEFs.

Original languageEnglish (US)
Pages (from-to)35654-35664
Number of pages11
JournalJournal of Biological Chemistry
Volume285
Issue number46
DOIs
StatePublished - Nov 12 2010

Fingerprint

p38 Mitogen-Activated Protein Kinases
Fibroblasts
Embryonic Structures
Acetylation
RNA Polymerase II
Transcription
Histones
Chromatin
Tumors
Exons
Neural Crest
Genes
Chromatin Immunoprecipitation
Chemical activation
Genetic Models
Embryonic Development
DNA
Neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Tgfβ signaling directly induces Arf promoter remodeling by a mechanism involving Smads 2/3 and p38 MAPK. / Zheng, Yanbin; Zhao, Yi D.; Gibbons, Melissa; Abramova, Tatiana; Chu, Patricia Y.; Ash, John D.; Cunningham, John M.; Skapek, Stephen X.

In: Journal of Biological Chemistry, Vol. 285, No. 46, 12.11.2010, p. 35654-35664.

Research output: Contribution to journalArticle

Zheng, Yanbin ; Zhao, Yi D. ; Gibbons, Melissa ; Abramova, Tatiana ; Chu, Patricia Y. ; Ash, John D. ; Cunningham, John M. ; Skapek, Stephen X. / Tgfβ signaling directly induces Arf promoter remodeling by a mechanism involving Smads 2/3 and p38 MAPK. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 46. pp. 35654-35664.
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AU - Ash, John D.

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