Screening of domain-specific target proteins of polo-like kinase 1: Construction and application of centrosome/kinetochore-specific targeting peptide

Jae Hoon Ji, Young Joo Jang

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Mammalian polo-like kinase 1 (Plk1) acts at various stages in early and late mitosis. Plk1 localizes at the centrosome and maintains this position through mitosis. Thereafter Plk1 moves to the kinetochore and midbody region, important sites during chromosome separation and cytokinesis. The catalytic domain of Plk1 is in the N-terminus region, whereas the non-catalytic region in the C-terminus of Plk1 has a conserved motif, named the Polobox. This motif is critical for Plk localization. EGFP proteins fused with the N-terminus and C-terminus of Plk1 localize in the nucleus and centrosomes, respectively. The core sequences of the polo-box (50 amino acids) also localize in Plk1 target organelles. To screen for domain-specific target proteins of Plk1, we constructed an N-terminal domain and a tandem repeat polo-box motif, and used them as templates in a yeast two-hybrid screen. The HeLa cell cDNA library indicated several proteins including the centrosome/kinetochore components or regulators, to be characterized as positive clones. Through in vitro protein binding analyses, we confirmed an interaction between these proteins and Plk1. The data reported from this study indicate that the N- and C-termini of Plk1 may function through recruitment and/or activation of domain-specific target proteins in dividing cells. Additionally, tandem repeats of the conserved core motif of the polo-box are sufficient for targeting and may be useful as a centrosome/kinetochore-specific targeting peptide.

Original languageEnglish (US)
Pages (from-to)709-716
Number of pages8
JournalJournal of Biochemistry and Molecular Biology
Volume39
Issue number6
DOIs
StatePublished - Nov 2006
Externally publishedYes

Keywords

  • Centrosome
  • Kinetochore
  • Polo-box motif
  • Polo-like kinase 1
  • Yeast two-hybrid screening

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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