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PMID: 12529437 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

CRM1/Ran-mediated nuclear export of p27(Kip1) involves a nuclear export signal and links p27 export and proteolysis.

Molecular biology of the cell ·Vol. 14 ·No. 1 ·2003-01-00 ·Pages 201-13

Connor MK, Kotchetkov R, Cariou S, Resch A, Lupetti R, Beniston RG, Melchior F, Hengst L, Slingerland JM

Abstract

We show that p27 localization is cell cycle regulated and we suggest that active CRM1/RanGTP-mediated nuclear export of p27 may be linked to cytoplasmic p27 proteolysis in early G1. p27 is nuclear in G0 and early G1 and appears transiently in the cytoplasm at the G1/S transition. Association of p27 with the exportin CRM1 was minimal in G0 and increased markedly during G1-to-S phase progression. Proteasome inhibition in mid-G1 did not impair nuclear import of p27, but led to accumulation of p27 in the cytoplasm, suggesting that export precedes degradation for at least part of the cellular p27 pool. p27-CRM1 binding and nuclear export were inhibited by S10A mutation but not by T187A mutation. A putative nuclear export sequence in p27 is identified whose mutation reduced p27-CRM1 interaction, nuclear export, and p27 degradation. Leptomycin B (LMB) did not inhibit p27-CRM1 binding, nor did it prevent p27 export in vitro or in heterokaryon assays. Prebinding of CRM1 to the HIV-1 Rev nuclear export sequence did not inhibit p27-CRM1 interaction, suggesting that p27 binds CRM1 at a non-LMB-sensitive motif. LMB increased total cellular p27 and may do so indirectly, through effects on other p27 regulatory proteins. These data suggest a model in which p27 undergoes active, CRM1-dependent nuclear export and cytoplasmic degradation in early G1. This would permit the incremental activation of cyclin E-Cdk2 leading to cyclin E-Cdk2-mediated T187 phosphorylation and p27 proteolysis in late G1 and S phase.

MeSH Terms
Active Transport, Cell Nucleus/physiology Animals Cell Cycle/physiology Cell Cycle Proteins/genetics,metabolism Cell Nucleus/metabolism Cyclin-Dependent Kinase Inhibitor p27 Cysteine Endopeptidases/metabolism HIV-1/metabolism Karyopherins/metabolism Mice Multienzyme Complexes/metabolism Mutation Proteasome Endopeptidase Complex Protein Sorting Signals/physiology Receptors, Cytoplasmic and Nuclear Tumor Suppressor Proteins/genetics,metabolism Viral Proteins/metabolism ran GTP-Binding Protein/metabolism
Chemicals
Cdkn1b protein, mouse Cell Cycle Proteins Karyopherins Multienzyme Complexes Protein Sorting Signals Receptors, Cytoplasmic and Nuclear Tumor Suppressor Proteins Viral Proteins exportin 1 protein Cyclin-Dependent Kinase Inhibitor p27 Cysteine Endopeptidases Proteasome Endopeptidase Complex ran GTP-Binding Protein
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Connor Michael K
Molecular and Cell Biology, Sunnybrook & Women's College Health Science Centre, 2075 Bayview Ave, Toronto, Ontario, Canada, M4N 3M5.
Kotchetkov Rouslan
Cariou Sandrine
Resch Ansgar
Lupetti Rafaella
Beniston Richard G
Melchior Frauke
Hengst Ludger
Slingerland Joyce M
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e02-06-0319
Published
2003-01-00
Pages
201-13
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC140238
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e02-06-0319 PMC Full Text View journal details
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