Interaction of Cep135 with a p50 Dynactin Subunit in Mammalian Centrosomes

Yumi Uetake, Yasuhiko Terada, Jurgita Matuliene, Ryoko Kuriyama

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Cep135 is a 135-kDa, coiled-coil centrosome protein important for microtubule organization in mammalian cells [Ohta et al., 2002: J. Cell Biol. 156: 87-99]. To identify Cep135-interacting molecules, we screened yeast two-hybrid libraries. One clone encoded dynamitin, a p50 dynactin subunit, which localized at the centrosome and has been shown to be involved in anchoring microtubules to centrosomes. The central domain of p50 binds to the C-terminal sequence of Cep135, this was further confirmed by immunoprecipitation and immunostaining of CHO cells co-expressing the binding domains for Cep135 and p50. Exogenous p50 lacking the Cep135-binding domain failed to locate at the centrosome, suggesting that Cep135 is required for initial targeting of the centrosome. Altered levels of Cep135 and p50 by RNAi and protein overexpression caused the release of endogenous partner molecules from centrosomes. This also resulted in dislocation of other centrosomal molecules, such as γ-tubulin and pericentrin, ultimately leading to disorganization of microtubule patterns. These results suggest that Cep135 and p50 play an important role in assembly and maintenance of functional microtubule-organizing centers. Cell Motil.

Original languageEnglish
Pages (from-to)53-66
Number of pages14
JournalCell Motility and the Cytoskeleton
Volume58
Issue number1
DOIs
Publication statusPublished - 2004 May
Externally publishedYes

Fingerprint

Centrosome
Microtubules
Microtubule-Organizing Center
Microtubule Proteins
CHO Cells
Tubulin
RNA Interference
Immunoprecipitation
Libraries
Dynactin Complex
Clone Cells
Yeasts
Maintenance
Proteins

Keywords

  • Centrosome
  • Cep135
  • Dynactin
  • Dynamitin
  • Microtubules
  • RNAi

ASJC Scopus subject areas

  • Cell Biology

Cite this

Interaction of Cep135 with a p50 Dynactin Subunit in Mammalian Centrosomes. / Uetake, Yumi; Terada, Yasuhiko; Matuliene, Jurgita; Kuriyama, Ryoko.

In: Cell Motility and the Cytoskeleton, Vol. 58, No. 1, 05.2004, p. 53-66.

Research output: Contribution to journalArticle

Uetake, Yumi ; Terada, Yasuhiko ; Matuliene, Jurgita ; Kuriyama, Ryoko. / Interaction of Cep135 with a p50 Dynactin Subunit in Mammalian Centrosomes. In: Cell Motility and the Cytoskeleton. 2004 ; Vol. 58, No. 1. pp. 53-66.
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