© The Rockefeller University Press,
0021-9525/1999//213 $5.00
The Journal of Cell Biology, Volume 144, Number 2,
, 1999 213-224
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 for both DNA replication and entry into mitosis. Consequently, most cyclin-dependent kinase (Cdk)/cyclin complexes are localized to the nucleus when active. Although our understanding of nuclear transport processes has been greatly enhanced by the recent identification of nuclear targeting sequences and soluble nuclear import factors with which they interact, the mechanisms used to target Cdk/cyclin complexes to the nucleus remain obscure; this is in part because these proteins lack obvious nuclear localization sequences. To elucidate the molecular mechanisms responsible for Cdk/cyclin transport, we examined nuclear import of fluorescent Cdk2/cyclin E and Cdc2/cyclin B1 complexes in digitonin-permeabilized mammalian cells and also examined potential physical interactions between these Cdks, cyclins, and soluble import factors. We found that the nuclear import machinery recognizes these Cdk/cyclin complexes through direct interactions with the cyclin component. Surprisingly, cyclins E and B1 are imported into nuclei via distinct mechanisms. Cyclin E behaves like a classical basic nuclear localization sequence–containing protein, binding to the
adaptor subunit of the importin-
/β heterodimer. In contrast, cyclin B1 is imported via a direct interaction with a site in the NH2 terminus of importin-β that is distinct from that used to bind importin-
.
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 Research Fund, Clare Hall Laboratories, Blanche Lane, South Mimms, Herts, EN6 3LD United Kingdom.

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