Specific Phosphorylation of Nucleophosmin on Thr199 by Cyclin-dependent Kinase 2-Cyclin E and Its Role in Centrosome Duplication

Yukari Tokuyama, Henning Horn, Kenji Kawamura, Pheruza Tarapore, Kenji Fukasawa

Research output: Contribution to journalArticle

177 Citations (Scopus)

Abstract

The kinase activity of cyclin-dependent kinase 2 (CDK2)-cyclin E is required for centrosomes to initiate duplication. We have recently found that nucleophosmin (NPM/B23), a phosphoprotein primarily found in nucleolus, associates with unduplicated centrosomes and is a direct substrate of CDK2-cyclin E in centrosome duplication. Upon phosphorylation by CDK2-cyclin E, NPM/B23 dissociates from centrosomes, which is a pre-requisite step for centrosomes to initiate duplication. Here, we identified that threonine 199 (Thr199) of NPM/B23 is the major phosphorylation target site of CDK2-cyclin E in vitro, and the same site is phosphorylated in vivo. NPM/T199A, a nonphosphorylatable NPM/B23 substitution mutant (Thr199 → Ala) acts as dominant negative when expressed in cells, resulting in specific inhibition of centrosome duplication. As expected, NPM/T199A remains associated with the centrosomes. These observations provide direct evidence that the CDK2-cyclin E-mediated phosphorylation on Thr199 determines association and dissociation of NPM/B23 to the centrosomes, which is a critical control for the centrosome to initiate duplication.

Original languageEnglish
Pages (from-to)21529-21537
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number24
DOIs
Publication statusPublished - 15 Jun 2001
Externally publishedYes

Fingerprint

Cyclin-Dependent Kinase 2
Cyclin E
Centrosome
Phosphorylation
Threonine
Phosphoproteins
Substitution reactions
Phosphotransferases
nucleophosmin
Association reactions
Substrates

ASJC Scopus subject areas

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

Cite this

Specific Phosphorylation of Nucleophosmin on Thr199 by Cyclin-dependent Kinase 2-Cyclin E and Its Role in Centrosome Duplication. / Tokuyama, Yukari; Horn, Henning; Kawamura, Kenji; Tarapore, Pheruza; Fukasawa, Kenji.

In: Journal of Biological Chemistry, Vol. 276, No. 24, 15.06.2001, p. 21529-21537.

Research output: Contribution to journalArticle

Tokuyama, Yukari ; Horn, Henning ; Kawamura, Kenji ; Tarapore, Pheruza ; Fukasawa, Kenji. / Specific Phosphorylation of Nucleophosmin on Thr199 by Cyclin-dependent Kinase 2-Cyclin E and Its Role in Centrosome Duplication. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 24. pp. 21529-21537.
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