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Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis.

Publication ,  Journal Article
Moore, JD; Kornbluth, S; Hunt, T
Published in: Molecular biology of the cell
December 2002

Cyclin-dependent kinase (Cdk)2/cyclin E is imported into nuclei assembled in Xenopus egg extracts by a pathway that requires importin-alpha and -beta. Here, we identify a basic nuclear localization sequence (NLS) in the N-terminus of Xenopus cyclin E. Mutation of the NLS eliminated nuclear accumulation of both cyclin E and Cdk2, and such versions of cyclin E were unable to trigger DNA replication. Addition of a heterologous NLS from SV40 large T antigen restored both nuclear targeting of Cdk2/cyclin E and DNA replication. We present evidence indicating that Cdk2/cyclin E complexes must become highly concentrated within nuclei to support replication and find that cyclin A can trigger replication at much lower intranuclear concentrations. We confirmed that depletion of endogenous cyclin E increases the concentration of cyclin B necessary to promote entry into mitosis. In contrast to its inability to promote DNA replication, cyclin E lacking its NLS was able to cooperate with cyclin B in promoting mitotic entry.

Duke Scholars

Published In

Molecular biology of the cell

DOI

EISSN

1939-4586

ISSN

1059-1524

Publication Date

December 2002

Volume

13

Issue

12

Start / End Page

4388 / 4400

Related Subject Headings

  • Xenopus Proteins
  • Xenopus
  • Sequence Homology, Amino Acid
  • Protein Structure, Tertiary
  • Protein Serine-Threonine Kinases
  • Plasmids
  • Nuclear Localization Signals
  • Mutation
  • Molecular Sequence Data
  • Mitosis
 

Citation

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MLA
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Moore, J. D., Kornbluth, S., & Hunt, T. (2002). Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis. Molecular Biology of the Cell, 13(12), 4388–4400. https://doi.org/10.1091/mbc.e02-07-0449
Moore, Jonathan D., Sally Kornbluth, and Tim Hunt. “Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis.Molecular Biology of the Cell 13, no. 12 (December 2002): 4388–4400. https://doi.org/10.1091/mbc.e02-07-0449.
Moore JD, Kornbluth S, Hunt T. Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis. Molecular biology of the cell. 2002 Dec;13(12):4388–400.
Moore, Jonathan D., et al. “Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis.Molecular Biology of the Cell, vol. 13, no. 12, Dec. 2002, pp. 4388–400. Epmc, doi:10.1091/mbc.e02-07-0449.
Moore JD, Kornbluth S, Hunt T. Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis. Molecular biology of the cell. 2002 Dec;13(12):4388–4400.

Published In

Molecular biology of the cell

DOI

EISSN

1939-4586

ISSN

1059-1524

Publication Date

December 2002

Volume

13

Issue

12

Start / End Page

4388 / 4400

Related Subject Headings

  • Xenopus Proteins
  • Xenopus
  • Sequence Homology, Amino Acid
  • Protein Structure, Tertiary
  • Protein Serine-Threonine Kinases
  • Plasmids
  • Nuclear Localization Signals
  • Mutation
  • Molecular Sequence Data
  • Mitosis