TY - JOUR
T1 - Minimal requirements for the nuclear localization of p27(Kip1), a cyclin-dependent kinase inhibitor
AU - Zeng, Ying
AU - Hirano, Katsuya
AU - Hirano, Mayumi
AU - Nishimura, Junji
AU - Kanaide, Hideo
N1 - Funding Information:
We thank Mr. Brian Quinn for comments and help with the manuscript. This study was supported in part by Grants-in-Aid for Scientific Research (Nos. 10557072, 11838013, and 11670687) and for Scientific Research on Priority Area (No. 12213103) from the Ministry of Education, Science, Sports, and Culture, Japan, by the Research Grant for Cardiovascular Diseases (11C-1) from the Ministry of Health and Welfare, Japan, and by grants from the Foundation for the Promotion of Clinical Medicine, the Suzuken Memorial Foundation, and KANZAWA Medical Research Foundation.
PY - 2000/7/21
Y1 - 2000/7/21
N2 - 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.
AB - 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.
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U2 - 10.1006/bbrc.2000.3098
DO - 10.1006/bbrc.2000.3098
M3 - Article
C2 - 10903892
AN - SCOPUS:0033913977
SN - 0006-291X
VL - 274
SP - 37
EP - 42
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -