Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1

K. Hirano, Mayumi Hirano, Y. Zeng, J. Nishimura, K. Hara, K. Muta, H. Nawata, H. Kanaide

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

p27Kip1 is an inhibitor of cyclin-dependent kinases. It has been implicated as having a role in the induction of growth arrest at the G1 phase of the cell cycle in response to anti-mitogenic signals such as cell contact and serum starvation. Proteasome-mediated degradation plays an important role in the rapid inactivation of p27Kip1, causing quiescent cells to re-enter the cell cycle. Although the existence of a second isoform has been suggested, no such isoform was isolated. Through screening of a cDNA library derived from growth-arrested confluent porcine endothelial cells, we obtained clones for a novel isoform of p27Kip1 in addition to the original isoform. The novel isoform differed from the original isoform at the C-terminus. The tissue-specific expression of the original and novel isoforms was demonstrated at the mRNA and protein levels. An in vitro degradation assay demonstrated this novel isoform to be resistant to proteasome-mediated destruction. The expression as a fusion protein with green fluorescent protein revealed this isoform to be targeted to the nucleus by a bipartite nuclear-localization signal with a C-terminal part different from that of the original isoform. The expression of the novel isoform caused the growth arrest of HeLa cells and an accumulation of cells in the G0/G1 phase, and this effect was similar to that seen with the original isoform. The present study suggests that the novel isoform functions as a negative regulator of the cell cycle, and may play a distinct role. The novel isoform was named p27Kip1R because of its resistance to degradation.

Original languageEnglish
Pages (from-to)51-57
Number of pages7
JournalBiochemical Journal
Volume353
Issue number1
DOIs
Publication statusPublished - Jan 1 2001

Fingerprint

Cloning
Organism Cloning
Protein Isoforms
Degradation
Cells
Cell Cycle
G1 Phase
Proteasome Endopeptidase Complex
Growth
Nuclear Localization Signals
Cell Cycle Resting Phase
Cyclin-Dependent Kinases
Endothelial cells
Starvation
Green Fluorescent Proteins
Gene Library
HeLa Cells
Assays
Screening
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hirano, K., Hirano, M., Zeng, Y., Nishimura, J., Hara, K., Muta, K., ... Kanaide, H. (2001). Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1. Biochemical Journal, 353(1), 51-57. https://doi.org/10.1042/bj3530051

Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1. / Hirano, K.; Hirano, Mayumi; Zeng, Y.; Nishimura, J.; Hara, K.; Muta, K.; Nawata, H.; Kanaide, H.

In: Biochemical Journal, Vol. 353, No. 1, 01.01.2001, p. 51-57.

Research output: Contribution to journalArticle

Hirano, K, Hirano, M, Zeng, Y, Nishimura, J, Hara, K, Muta, K, Nawata, H & Kanaide, H 2001, 'Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1', Biochemical Journal, vol. 353, no. 1, pp. 51-57. https://doi.org/10.1042/bj3530051
Hirano, K. ; Hirano, Mayumi ; Zeng, Y. ; Nishimura, J. ; Hara, K. ; Muta, K. ; Nawata, H. ; Kanaide, H. / Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1. In: Biochemical Journal. 2001 ; Vol. 353, No. 1. pp. 51-57.
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