Minimal requirements for the nuclear localization of p27(Kip1), a cyclin-dependent kinase inhibitor

Ying Zeng, Katsuya Hirano, Mayumi Hirano, Junji Nishimura, Hideo Kanaide

    Research output: Contribution to journalArticle

    50 Citations (Scopus)

    Abstract

    p27(Kip1) is a cyclin-dependent kinase inhibitor, and its nuclear localization is a prerequisite for it to function as a cell cycle regulator. In the present study, the minimal requirement for the nuclear localization signal (NLS) of p27(Kip1) was determined by analyzing the localization of various mutants of p27(Kip1) tagged with green fluorescent protein (GFP) in HeLa cells and porcine aortic endothelial cells. Wild-type p27(Kip1) exclusively localized into nucleus, while GFP alone localized in both cytosol and nucleus. A comparison of various truncation mutants revealed residues 153-166 to be the minimal region necessary for nuclear localization. However, a fusion of this region to GFP showed cytoplasmic retention in addition to nuclear localization, thus suggesting that some extension flanking this region is required to achieve a full function of NLS. The site-directed mutation of the full-length p27(Kip1) therefore showed that four basic residues (K153, R154, K165, R166), especially R166, play a critical role in the nuclear localization of p27(Kip1). (C) 2000 Academic Press.

    Original languageEnglish
    Pages (from-to)37-42
    Number of pages6
    JournalBiochemical and Biophysical Research Communications
    Volume274
    Issue number1
    DOIs
    Publication statusPublished - Jul 21 2000

    All Science Journal Classification (ASJC) codes

    • Biophysics
    • Biochemistry
    • Molecular Biology
    • Cell Biology

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