Polo-box motif targets a centrosome regulator, RanGTPase

Young Joo Jang, Jae Hoon Ji, Ji Hee Ahn, Kwang Lae Hoe, Misun Won, Dong Soo Im, Suhn Kee Chae, Sukgil Song, Hyang Sook Yoo

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Mammalian polo-like kinase (Plk) acts at various stages in early and late mitosis. Plk1 localizes in the centrosome, the central spindle, the midbody as well as the kinetochore. The non-catalytic region in the C-terminus of Plk1 has conserved sequence motifs, named polo-boxes. These motifs are important for Plk localization. GFP protein fused with the core sequences of polo-box (50 amino acids) localized Plk to target organelles. We screened for Plk interacting proteins by constructing a tandem repeat of the polo-box motif, and used it as bait in the two-hybrid system with HeLa cell cDNA library. RanGTPase was detected as a positive clone. Through in vitro and in vivo protein binding analysis in synchronized cells by thymidine block and by nocodazole treatment, we confirmed the interaction between endogenous Ran and Plk1. We showed that endogenous Ran and Plk1 proteins were co-localized to centrosomes, which is a major target organelle of endogenous Plk1, in early mitotic cells by immunofluorescence. Finally, we demonstrated that Plk1 phosphorylated RanBPM, a Ran-binding protein in microtubule organizing center, through the interaction with Ran. These data suggested that the core motif of polo-box is sufficient for Plk1-targeting, and that Plk1 may play roles in centrosome through recruitment and/or activation of Ran/RanBPM proteins.

Original languageEnglish (US)
Pages (from-to)257-264
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume325
Issue number1
DOIs
StatePublished - Dec 3 2004
Externally publishedYes

Keywords

  • Centrosome
  • Polo-box
  • Polo-like kinase
  • RanBPM
  • RanGTPase

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry
  • Cell Biology

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