Nuclear Import of Cdk/Cyclin Complexes: Identification of Distinct Mechanisms for Import of Cdk2/Cyclin E and Cdc2/Cyclin B1

doi:10.1083/jcb.144.2.213

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© The Rockefeller University Press, 0021-9525/1999//213 $5.00
The Journal of Cell Biology, Volume 144, Number 2, , 1999 213-224

Regular Articles

Nuclear Import of Cdk/Cyclin Complexes: Identification of Distinct Mechanisms for Import of Cdk2/Cyclin E and Cdc2/Cyclin B1

Jonathan D. Moore, Jing Yang, Ray Truant^*, and Sally Kornbluth

Department of Pharmacology and Cancer Biology, and ^* Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710

Reversible phosphorylation of nuclear proteins is required forboth DNA replication and entry into mitosis. Consequently,most cyclin-dependent kinase (Cdk)/cyclin complexes are localizedto the nucleus when active. Although our understanding of nucleartransport processes has been greatly enhanced by the recentidentification of nuclear targeting sequences and soluble nuclearimport factors with which they interact, the mechanisms usedto target Cdk/cyclin complexes to the nucleus remain obscure;this is in part because these proteins lack obvious nuclearlocalization sequences. To elucidate the molecular mechanisms responsible for Cdk/cyclin transport, we examined nuclearimport of fluorescent Cdk2/cyclin E and Cdc2/cyclin B1 complexesin digitonin-permeabilized mammalian cells and also examinedpotential physical interactions between these Cdks, cyclins,and soluble import factors. We found that the nuclear importmachinery recognizes these Cdk/cyclin complexes through directinteractions with the cyclin component. Surprisingly, cyclinsE and B1 are imported into nuclei via distinct mechanisms. CyclinE behaves like a classical basic nuclear localization sequence–containingprotein, binding to the ${alpha}$ adaptor subunit of the importin- ${alpha}$ /β heterodimer. In contrast, cyclin B1 is imported via a directinteraction with a site in the NH₂ terminus of importin-βthat is distinct from that used to bind importin- ${alpha}$ .

Key Words: importins • cyclins • Cdk • nuclear import • mitosis

Abbreviations used in this paper: Cdk, cyclin-dependent kinase; IBB, importin-β binding; NES, nuclear export sequence; NLS, nuclear localization sequence.

Jonathan D. Moore's current address is Imperial Cancer ResearchFund, Clare Hall Laboratories, Blanche Lane, South Mimms, Herts,EN6 3LD United Kingdom.

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