Transforming growth factor β facilitates β-TrCP-mediated degradation of Cdc25A in a Smad3-dependent manner

Dipankar Ray, Yasuhisa Terao, Dipali Nimbalkar, Li Hao Chu, Maddalena Donzelli, Tateki Tsutsui, Xianghong Zou, Asish K. Ghosh, John Varga, Giulio F. Draetta, Hiroaki Kiyokawa*

*Corresponding author for this work

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Ubiquitin-dependent degradation of Cdc25A is a major mechanism for damage-induced S-phase checkpoint. Two ubiquitin ligases, the Skp1-cullin-β-TrCP (SCFβ-TrCP) complex and the anaphase-promoting complex (APCCdh1), are involved in Cdc25A degradation. Here we demonstrate that the transforming growth factor β (TGF-β)-Smad3 pathway promotes SCFβ-TrCP-mediated Cdc25A ubiquitination. Cells treated with TGF-β, as well as cells transfected with Smad3 or a constitutively active type I TGF-β receptor, exhibit increased ubiquitination and markedly shortened half-lives of Cdc25A. Furthermore, Cdc25A is stabilized in cells transfected with Smad3 small interfering RNA (siRNA) and cells from Smad3-null mice. TGF-β-induced ubiquitination is associated with Cdc25A phosphorylation at the β-TrCP docking site (DS82G motif) and physical association of Cdc25A with Smad3 and β-TrCP. Cdc25A mutant proteins deficient in DS82G phosphorylation are resistant to TGF-β-Smad3-induced degradation, whereas a Cdc25A mutant protein defective in APCCdh1 recognition undergoes efficient degradation. Smad3 siRNA inhibits β-TrCP-Cdc25A interaction and Cdc25A degradation in response to TGF-β. β-TrCP2 siRNA also inhibits Smad3-induced Cdc25A degradation. In contrast, Cdh1 siRNA had no effect on Cdc25A down-regulation by Smad3. These data suggest that Smad3 plays a key role in the regulation of Cdc25A ubiquitination by SCFβ-TrCP and that Cdc25A stabilization observed in various cancers could be associated with defects in the TGF-β-Smad3 pathway.

Original languageEnglish (US)
Pages (from-to)3338-3347
Number of pages10
JournalMolecular and cellular biology
Volume25
Issue number8
DOIs
StatePublished - Apr 1 2005

Fingerprint

Transforming Growth Factors
Ubiquitination
Small Interfering RNA
Mutant Proteins
Ubiquitin
S Phase Cell Cycle Checkpoints
Cullin Proteins
Phosphorylation
Anaphase-Promoting Complex-Cyclosome
Null Lymphocytes
Growth Factor Receptors
Ligases
Down-Regulation
Neoplasms

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Ray, Dipankar ; Terao, Yasuhisa ; Nimbalkar, Dipali ; Chu, Li Hao ; Donzelli, Maddalena ; Tsutsui, Tateki ; Zou, Xianghong ; Ghosh, Asish K. ; Varga, John ; Draetta, Giulio F. ; Kiyokawa, Hiroaki. / Transforming growth factor β facilitates β-TrCP-mediated degradation of Cdc25A in a Smad3-dependent manner. In: Molecular and cellular biology. 2005 ; Vol. 25, No. 8. pp. 3338-3347.
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abstract = "Ubiquitin-dependent degradation of Cdc25A is a major mechanism for damage-induced S-phase checkpoint. Two ubiquitin ligases, the Skp1-cullin-β-TrCP (SCFβ-TrCP) complex and the anaphase-promoting complex (APCCdh1), are involved in Cdc25A degradation. Here we demonstrate that the transforming growth factor β (TGF-β)-Smad3 pathway promotes SCFβ-TrCP-mediated Cdc25A ubiquitination. Cells treated with TGF-β, as well as cells transfected with Smad3 or a constitutively active type I TGF-β receptor, exhibit increased ubiquitination and markedly shortened half-lives of Cdc25A. Furthermore, Cdc25A is stabilized in cells transfected with Smad3 small interfering RNA (siRNA) and cells from Smad3-null mice. TGF-β-induced ubiquitination is associated with Cdc25A phosphorylation at the β-TrCP docking site (DS82G motif) and physical association of Cdc25A with Smad3 and β-TrCP. Cdc25A mutant proteins deficient in DS82G phosphorylation are resistant to TGF-β-Smad3-induced degradation, whereas a Cdc25A mutant protein defective in APCCdh1 recognition undergoes efficient degradation. Smad3 siRNA inhibits β-TrCP-Cdc25A interaction and Cdc25A degradation in response to TGF-β. β-TrCP2 siRNA also inhibits Smad3-induced Cdc25A degradation. In contrast, Cdh1 siRNA had no effect on Cdc25A down-regulation by Smad3. These data suggest that Smad3 plays a key role in the regulation of Cdc25A ubiquitination by SCFβ-TrCP and that Cdc25A stabilization observed in various cancers could be associated with defects in the TGF-β-Smad3 pathway.",
author = "Dipankar Ray and Yasuhisa Terao and Dipali Nimbalkar and Chu, {Li Hao} and Maddalena Donzelli and Tateki Tsutsui and Xianghong Zou and Ghosh, {Asish K.} and John Varga and Draetta, {Giulio F.} and Hiroaki Kiyokawa",
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Transforming growth factor β facilitates β-TrCP-mediated degradation of Cdc25A in a Smad3-dependent manner. / Ray, Dipankar; Terao, Yasuhisa; Nimbalkar, Dipali; Chu, Li Hao; Donzelli, Maddalena; Tsutsui, Tateki; Zou, Xianghong; Ghosh, Asish K.; Varga, John; Draetta, Giulio F.; Kiyokawa, Hiroaki.

In: Molecular and cellular biology, Vol. 25, No. 8, 01.04.2005, p. 3338-3347.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Transforming growth factor β facilitates β-TrCP-mediated degradation of Cdc25A in a Smad3-dependent manner

AU - Ray, Dipankar

AU - Terao, Yasuhisa

AU - Nimbalkar, Dipali

AU - Chu, Li Hao

AU - Donzelli, Maddalena

AU - Tsutsui, Tateki

AU - Zou, Xianghong

AU - Ghosh, Asish K.

AU - Varga, John

AU - Draetta, Giulio F.

AU - Kiyokawa, Hiroaki

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N2 - Ubiquitin-dependent degradation of Cdc25A is a major mechanism for damage-induced S-phase checkpoint. Two ubiquitin ligases, the Skp1-cullin-β-TrCP (SCFβ-TrCP) complex and the anaphase-promoting complex (APCCdh1), are involved in Cdc25A degradation. Here we demonstrate that the transforming growth factor β (TGF-β)-Smad3 pathway promotes SCFβ-TrCP-mediated Cdc25A ubiquitination. Cells treated with TGF-β, as well as cells transfected with Smad3 or a constitutively active type I TGF-β receptor, exhibit increased ubiquitination and markedly shortened half-lives of Cdc25A. Furthermore, Cdc25A is stabilized in cells transfected with Smad3 small interfering RNA (siRNA) and cells from Smad3-null mice. TGF-β-induced ubiquitination is associated with Cdc25A phosphorylation at the β-TrCP docking site (DS82G motif) and physical association of Cdc25A with Smad3 and β-TrCP. Cdc25A mutant proteins deficient in DS82G phosphorylation are resistant to TGF-β-Smad3-induced degradation, whereas a Cdc25A mutant protein defective in APCCdh1 recognition undergoes efficient degradation. Smad3 siRNA inhibits β-TrCP-Cdc25A interaction and Cdc25A degradation in response to TGF-β. β-TrCP2 siRNA also inhibits Smad3-induced Cdc25A degradation. In contrast, Cdh1 siRNA had no effect on Cdc25A down-regulation by Smad3. These data suggest that Smad3 plays a key role in the regulation of Cdc25A ubiquitination by SCFβ-TrCP and that Cdc25A stabilization observed in various cancers could be associated with defects in the TGF-β-Smad3 pathway.

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