Involvement of GSK-3β and DYRK1B in differentiation-inducing factor-3-induced phosphorylation of cyclin D1 in HeLa cells

Fumi Takahashi, Jun Mori, Etsuko Matsuzaki, Yutaka Watanabe, Masato Hirata, Yoshikazu Miwa, Sachio Morimoto, Toshiyuki Sasaguri

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Differentiation-inducing factors (DIFs) are putative morphogens that induce cell differentiation in Dictyostelium discoideum. We previously reported that DIF-3 activates glycogen synthase kinase-3β (GSK-3β), resulting in the degradation of cyclin D1 in HeLa cells. In this study, we investigated the effect of DIF-3 on cyclin D1 mutants (R29Q, L32A, T286A, T288A, and T286A/T288A) to clarify the precise mechanisms by which DIF-3 degrades cyclin D1 in HeLa cells. We revealed that T286A, T288A, and T286A/T288A mutants were resistant to DIF-3-induced degradation compared with wild-type cyclin D1, indicating that the phosphorylation of Thr286 and Thr288 were critical for cyclin D1 degradation induced by DIF-3. Indeed, DIF-3 markedly elevated the phosphorylation level of cyclin D1, and mutations introduced to Thr 286 and/or Thr288 prevented the phosphorylation induced by DIF-3. Depletion of endogenous GSK-3β and dual-specificity tyrosine phosphorylation regulated kinase 1B (DYRK1B) by RNA interference attenuated the DIF-3-induced cyclin D1 phosphorylation and degradation. The effect of DIF-3 on DYRK1B activity was examined and we found that DIF-3 also activated this kinase. Further, we found that not only GSK-3β but also DYRK1B modulates cyclin D1 subcellular localization by the phosphorylation of Thr288. These results suggest that DIF-3 induces degradation of cyclin D1 through the GSK-3β- and DYRK1B-mediated threonine phosphorylation in HeLa cells.

Original languageEnglish
Pages (from-to)38489-38497
Number of pages9
JournalJournal of Biological Chemistry
Volume281
Issue number50
DOIs
Publication statusPublished - Dec 15 2006

Fingerprint

Glycogen Synthase Kinase 3
Phosphorylation
Cyclin D1
HeLa Cells
Degradation
Dyrk kinase
1-((3,5-dichloro)-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone
Dictyostelium
Threonine
RNA Interference
Cell Differentiation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Involvement of GSK-3β and DYRK1B in differentiation-inducing factor-3-induced phosphorylation of cyclin D1 in HeLa cells. / Takahashi, Fumi; Mori, Jun; Matsuzaki, Etsuko; Watanabe, Yutaka; Hirata, Masato; Miwa, Yoshikazu; Morimoto, Sachio; Sasaguri, Toshiyuki.

In: Journal of Biological Chemistry, Vol. 281, No. 50, 15.12.2006, p. 38489-38497.

Research output: Contribution to journalArticle

Takahashi, Fumi ; Mori, Jun ; Matsuzaki, Etsuko ; Watanabe, Yutaka ; Hirata, Masato ; Miwa, Yoshikazu ; Morimoto, Sachio ; Sasaguri, Toshiyuki. / Involvement of GSK-3β and DYRK1B in differentiation-inducing factor-3-induced phosphorylation of cyclin D1 in HeLa cells. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 50. pp. 38489-38497.
@article{dd82dc9d6e264775866438364da62b69,
title = "Involvement of GSK-3β and DYRK1B in differentiation-inducing factor-3-induced phosphorylation of cyclin D1 in HeLa cells",
abstract = "Differentiation-inducing factors (DIFs) are putative morphogens that induce cell differentiation in Dictyostelium discoideum. We previously reported that DIF-3 activates glycogen synthase kinase-3β (GSK-3β), resulting in the degradation of cyclin D1 in HeLa cells. In this study, we investigated the effect of DIF-3 on cyclin D1 mutants (R29Q, L32A, T286A, T288A, and T286A/T288A) to clarify the precise mechanisms by which DIF-3 degrades cyclin D1 in HeLa cells. We revealed that T286A, T288A, and T286A/T288A mutants were resistant to DIF-3-induced degradation compared with wild-type cyclin D1, indicating that the phosphorylation of Thr286 and Thr288 were critical for cyclin D1 degradation induced by DIF-3. Indeed, DIF-3 markedly elevated the phosphorylation level of cyclin D1, and mutations introduced to Thr 286 and/or Thr288 prevented the phosphorylation induced by DIF-3. Depletion of endogenous GSK-3β and dual-specificity tyrosine phosphorylation regulated kinase 1B (DYRK1B) by RNA interference attenuated the DIF-3-induced cyclin D1 phosphorylation and degradation. The effect of DIF-3 on DYRK1B activity was examined and we found that DIF-3 also activated this kinase. Further, we found that not only GSK-3β but also DYRK1B modulates cyclin D1 subcellular localization by the phosphorylation of Thr288. These results suggest that DIF-3 induces degradation of cyclin D1 through the GSK-3β- and DYRK1B-mediated threonine phosphorylation in HeLa cells.",
author = "Fumi Takahashi and Jun Mori and Etsuko Matsuzaki and Yutaka Watanabe and Masato Hirata and Yoshikazu Miwa and Sachio Morimoto and Toshiyuki Sasaguri",
year = "2006",
month = "12",
day = "15",
doi = "10.1074/jbc.M605205200",
language = "English",
volume = "281",
pages = "38489--38497",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "50",

}

TY - JOUR

T1 - Involvement of GSK-3β and DYRK1B in differentiation-inducing factor-3-induced phosphorylation of cyclin D1 in HeLa cells

AU - Takahashi, Fumi

AU - Mori, Jun

AU - Matsuzaki, Etsuko

AU - Watanabe, Yutaka

AU - Hirata, Masato

AU - Miwa, Yoshikazu

AU - Morimoto, Sachio

AU - Sasaguri, Toshiyuki

PY - 2006/12/15

Y1 - 2006/12/15

N2 - Differentiation-inducing factors (DIFs) are putative morphogens that induce cell differentiation in Dictyostelium discoideum. We previously reported that DIF-3 activates glycogen synthase kinase-3β (GSK-3β), resulting in the degradation of cyclin D1 in HeLa cells. In this study, we investigated the effect of DIF-3 on cyclin D1 mutants (R29Q, L32A, T286A, T288A, and T286A/T288A) to clarify the precise mechanisms by which DIF-3 degrades cyclin D1 in HeLa cells. We revealed that T286A, T288A, and T286A/T288A mutants were resistant to DIF-3-induced degradation compared with wild-type cyclin D1, indicating that the phosphorylation of Thr286 and Thr288 were critical for cyclin D1 degradation induced by DIF-3. Indeed, DIF-3 markedly elevated the phosphorylation level of cyclin D1, and mutations introduced to Thr 286 and/or Thr288 prevented the phosphorylation induced by DIF-3. Depletion of endogenous GSK-3β and dual-specificity tyrosine phosphorylation regulated kinase 1B (DYRK1B) by RNA interference attenuated the DIF-3-induced cyclin D1 phosphorylation and degradation. The effect of DIF-3 on DYRK1B activity was examined and we found that DIF-3 also activated this kinase. Further, we found that not only GSK-3β but also DYRK1B modulates cyclin D1 subcellular localization by the phosphorylation of Thr288. These results suggest that DIF-3 induces degradation of cyclin D1 through the GSK-3β- and DYRK1B-mediated threonine phosphorylation in HeLa cells.

AB - Differentiation-inducing factors (DIFs) are putative morphogens that induce cell differentiation in Dictyostelium discoideum. We previously reported that DIF-3 activates glycogen synthase kinase-3β (GSK-3β), resulting in the degradation of cyclin D1 in HeLa cells. In this study, we investigated the effect of DIF-3 on cyclin D1 mutants (R29Q, L32A, T286A, T288A, and T286A/T288A) to clarify the precise mechanisms by which DIF-3 degrades cyclin D1 in HeLa cells. We revealed that T286A, T288A, and T286A/T288A mutants were resistant to DIF-3-induced degradation compared with wild-type cyclin D1, indicating that the phosphorylation of Thr286 and Thr288 were critical for cyclin D1 degradation induced by DIF-3. Indeed, DIF-3 markedly elevated the phosphorylation level of cyclin D1, and mutations introduced to Thr 286 and/or Thr288 prevented the phosphorylation induced by DIF-3. Depletion of endogenous GSK-3β and dual-specificity tyrosine phosphorylation regulated kinase 1B (DYRK1B) by RNA interference attenuated the DIF-3-induced cyclin D1 phosphorylation and degradation. The effect of DIF-3 on DYRK1B activity was examined and we found that DIF-3 also activated this kinase. Further, we found that not only GSK-3β but also DYRK1B modulates cyclin D1 subcellular localization by the phosphorylation of Thr288. These results suggest that DIF-3 induces degradation of cyclin D1 through the GSK-3β- and DYRK1B-mediated threonine phosphorylation in HeLa cells.

UR - http://www.scopus.com/inward/record.url?scp=33845985316&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845985316&partnerID=8YFLogxK

U2 - 10.1074/jbc.M605205200

DO - 10.1074/jbc.M605205200

M3 - Article

C2 - 17046823

AN - SCOPUS:33845985316

VL - 281

SP - 38489

EP - 38497

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 50

ER -