Advertisement
Journal Article
The Saccharomyces cerevisiae cyclin Clb2p is targeted to multiple subcellular locations by cis- and trans-acting determinants
J.K. Hood, W.W. Hwang, P.A. Silver
Journal of Cell Science 2001 114: 589-597;

Summary

The cyclin-dependent kinase Cdc28p associates with the cyclin Clb2p to induce mitosis in the yeast Saccharomyces cerevisiae. Several cell cycle regulatory proteins have been shown to require specific nuclear transport events to exert their regulatory functions. Therefore, we investigated the subcellular localization of wild-type Clb2p and several mutant versions of the protein using green fluorescent protein (GFP) fusion constructs. Wild-type Clb2p is primarily nuclear at all points of the cell. A point mutation in a potential leucine-rich nuclear export signal (NES) enhances the nuclear localization of the protein, and delta-yrb2 cells exhibit an apparent Clb2p nuclear export defect. Clb2p contains a bipartite nuclear localization signal (NLS), and its nuclear localization requires the alpha and beta importins (Srp1p and Kap95p), as well as the yeast Ran GTPase and its regulators. Deletion of the Clb2p NLS causes increased cytoplasmic localization of the protein, as well as accumulation at the bud neck. These data indicate that Clb2p exists in multiple places in the yeast cell, possibly allowing Cdc28p to locally phosphorylate substrates at distinct subcellular sites.

REFERENCES

    1. Altman, R. and
    2. Kellogg, D.
    (1997). Control of mitotic events by Nap1 and the Gin4 kinase. J. Cell Biol 138, 119–130
    OpenUrlAbstract/FREE Full Text
    1. Amon, A.,
    2. Irniger, S. and
    3. Nasmyth, K.
    (1994). Closing the cell cycle circle in yeast: G2 cyclin proteolysis initiated at mitosis persists until the activation of G1 cyclins in the next cell cycle. Cell 77, 1037–1050
    OpenUrlCrossRefPubMedWeb of Science
    1. Amon, A.,
    2. Tyers, M.,
    3. Futcher, B. and
    4. Nasmyth, K.
    (1993). Mechanisms that help the yeast cell cycle clock tick: G2 cyclins transcriptionally activate G2 cyclins and repress G1 cyclins. Cell 74, 993–1007
    OpenUrlCrossRefPubMedWeb of Science
    1. Brandeis, M. and
    2. Hunt, T.
    (1996). The proteolysis of mitotic cyclins in mammalian cells persists from the end of mitosis until the onset of S phase. EMBO J 15, 5280–5289
    OpenUrlPubMedWeb of Science
    1. Corbett, A. H.,
    2. Koepp, D. M.,
    3. Lee, M. S.,
    4. Schlenstedt, G.,
    5. Hopper, A. K. and
    6. Silver, P. A.
    (1995). Rna1p, a Ran/TC4 GTPase activating protein, is required for nuclear import. J. Cell Biol 130, 1017–1026
    OpenUrlAbstract/FREE Full Text
    1. Fitch, I.,
    2. Dahmann, C.,
    3. Surana, U.,
    4. Amon, A.,
    5. Nasmyth, K.,
    6. Goetsch, L.,
    7. Byers, B. and
    8. Futcher, B.
    (1992). Characterization of four B-type cyclin genes of the budding yeast Saccharomyces cerevisiae. Mol. Biol. Cell 3, 805–818
    OpenUrlAbstract/FREE Full Text
    1. Ford, S. K. and
    2. Pringle, J. R.
    (1991). Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC11 gene product and the timing of events at the budding site. Dev. Genet 12, 281–292
    OpenUrlCrossRefPubMedWeb of Science
    1. Fridell, R. A.,
    2. Bogerd, H. P. and
    3. Cullen, B. R.
    (1996). Nuclear export of late HIV-1 mRNAs occurs via a cellular protein export pathway. Proc. Natl. Acad. Sci. USA 93, 4421–4424
    OpenUrlAbstract/FREE Full Text
    1. Görlich, D. and
    2. Kutay, U.
    (1999). Transport between the cell nucleus and the cytoplasm. Annu. Rev. Cell Dev. Biol 15, 607–660
    OpenUrlCrossRefPubMedWeb of Science
    1. Haarer, B. K. and
    2. Pringle, J. R.
    (1987). Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck. Mol. Cell. Biol 7, 3678–3687
    OpenUrlAbstract/FREE Full Text
    1. Hagting, A.,
    2. Karlsson, C.,
    3. Clute, P.,
    4. Jackman, M. and
    5. Pines, J.
    (1998). MPF localization is controlled by nuclear export. EMBO J 17, 4127–4138
    OpenUrlAbstract
    1. Hood, J. K. and
    2. Silver, P. A.
    (1998). Cse1p is required for export of Srp1p/importin-from the nucleus in Saccharomyces cerevisiae. J. Biol. Chem 273, 35142–35146
    OpenUrlAbstract/FREE Full Text
    1. Kellogg, D. R.,
    2. Kikuchi, A.,
    3. Fujii-Nakata, T.,
    4. Turck, C. W. and
    5. Murray, A. W.
    (1995). Members of the NAP/SET family of proteins interact specifically with B-type cyclins. J. Cell Biol 130, 661–673
    OpenUrlAbstract/FREE Full Text
    1. Kim, H. B.,
    2. Haarer, B. K. and
    3. Pringle, J. R.
    (1991). Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC3 gene product and the timing of events at the budding site. J. Cell Biol 112, 535–544
    OpenUrlAbstract/FREE Full Text
    1. King, R. W.,
    2. Peters, J. M.,
    3. Tugendreich, S.,
    4. Rolfe, M.,
    5. Hieter, P. and
    6. Kirschner, M. W.
    (1995). A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B. Cell 81, 279–288
    OpenUrlCrossRefPubMedWeb of Science
    1. Kong, M.,
    2. Barnes, E. A.,
    3. Ollendorff, V. and
    4. Donoghue, D. J.
    (2000). Cyclin F regulates the nuclear localization of cyclin B1 through a cyclin-cyclin interaction. EMBO J 19, 1378–1388
    OpenUrlAbstract/FREE Full Text
    1. Lew, D. J. and
    2. Reed, S. I.
    (1993). Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins. J. Cell Biol 120, 1305–1320
    OpenUrlAbstract/FREE Full Text
    1. Loeb, J. D. L.,
    2. Schlenstedt, G.,
    3. Pellman, D.,
    4. Kornitzer, D.,
    5. Silver, P. A. and
    6. Fink, G.
    (1995). The yeast nuclear import receptor is required for mitosis. Proc. Natl. Acad. Sci. USA 92, 7647–7651
    OpenUrlAbstract/FREE Full Text
    1. Meyer, B. E. and
    2. Malim, M. H.
    (1994). The HIV-1 Rev trans-activtor shuttles between the nucleus and the cytoplasm. Genes Dev 8, 1538–1547
    OpenUrlAbstract/FREE Full Text
    1. Moore, J. D.,
    2. Yang, J.,
    3. Truant, R. and
    4. Kornbluth, S.
    (1999). Nuclear import of Cdk/cyclin complexes: identification of distinct mechanisms for import of Cdk2/cyclin E and Cdc2/cyclin B1. J. Cell Biol 144, 213–224
    OpenUrlAbstract/FREE Full Text
    1. Murphy, R. and
    2. Wente, S. R.
    (1996). An RNA-export mediator with an essential nuclear export signal. Nature 383, 357–360
    OpenUrlCrossRefPubMedWeb of Science
    1. Nasmyth, K.
    (1993). Control of the yeast cell cycle by the Cdc28 protein kinase. Curr. Opin. Cell Biol 5, 166–179
    OpenUrlCrossRefPubMed
    1. Noguchi, E.,
    2. Saitoh, Y.,
    3. Sazer, S. and
    4. Nishimoto, T.
    (1999). Disruption of the YRB2 gene retards nuclear protein export, causing a profound mitotic delay and can be rescued by overexpression of XPO1/CRM1. J. Biochem. (Tokyo) 125, 574–585
    OpenUrlAbstract/FREE Full Text
    1. Pines, J.
    (1995). Cyclins and cyclin-dependent kinases: a biochemical view. Biochem. J 308, 697–711
    OpenUrlFREE Full Text
    1. Pines, J. and
    2. Hunter, T.
    (1991). Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport. J. Cell Biol 115, 1–17
    OpenUrlAbstract/FREE Full Text
    1. Richardson, H.,
    2. Lew, D. J.,
    3. Henze, M.,
    4. Sugimoto, K. and
    5. Reed, S. I.
    (1992). Cyclin-B homologs in Saccharomyces cerevisiae function in S phase and in G2. Genes Dev 6, 2021–2034
    OpenUrlAbstract/FREE Full Text
    1. Schlenstedt, G.,
    2. Wong, D. H.,
    3. Koepp, D. M. and
    4. Silver, P. A.
    (1995). Mutants in a yeast Ran binding protein are defective in nuclear transport. EMBO J 14, 5367–5378
    OpenUrlPubMedWeb of Science
    1. Seedorf, M.,
    2. Damelin, M.,
    3. Kahana, J.,
    4. Taura, T. and
    5. Silver, P. A.
    (1999). Interactions between a nuclear transporter and a subset of nuclear porecomplex proteins depend on Ran GTPase. Mol. Cell Biol 19, 1547–1557
    OpenUrlAbstract/FREE Full Text
    1. Seedorf, M. and
    2. Silver, P. A.
    (1997). Importin/karyopherin protein family members required for mRNA export from the nucleus. Proc. Natl. Acad. Sci. USA 94, 8590–8595
    OpenUrlAbstract/FREE Full Text
    1. Sudakin, V.,
    2. Ganoth, D.,
    3. Dahan, A.,
    4. Heller, H.,
    5. Hershko, J.,
    6. Luca, F. C.,
    7. Ruderman, J. V. and
    8. Hershko, A.
    (1995). The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. Mol. Biol. Cell 6, 185–197
    OpenUrlAbstract/FREE Full Text
    1. Surana, U.,
    2. Amon, A.,
    3. Dowzer, C.,
    4. McGrew, J.,
    5. Byers, B. and
    6. Nasmyth, K.
    (1993). Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J 12, 1969–1978
    OpenUrlPubMedWeb of Science
    1. Taura, T.,
    2. Krebber, H. and
    3. Silver, P. A.
    (1998). A member of the Ran-binding protein family, Yrb2p, is involved in nuclear protein export. Proc. Natl. Acad. Sci. USA 95, 7427–7432
    OpenUrlAbstract/FREE Full Text
    1. Toyoshima, F.,
    2. Moriguchi, T.,
    3. Wada, A.,
    4. Fukuda, M. and
    5. Nishida, E.
    (1998). Nuclear export of cyclin B1 and its possible role in the DNA damage-induced G2checkpoint. EMBO J 17, 2728–2735
    OpenUrlAbstract
    1. Wen, W.,
    2. Meinkoth, J. L.,
    3. Tsien, R. Y. and
    4. Taylor, S. S.
    (1995). Identification of a signal for rapid export of proteins from the nucleus. Cell 82, 463–473
    OpenUrlCrossRefPubMedWeb of Science
    1. Winston, F.,
    2. Dollard, C.,
    3. Dollard, S.L. and
    4. Dollard, R.-H.
    (1995). Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast 11, 53–55
    OpenUrlCrossRefPubMedWeb of Science
    1. Wong, D. H.,
    2. Corbett, A. H.,
    3. Kent, H. M.,
    4. Stewart, M. and
    5. Silver, P. A.
    (1997). Interaction between the small GTPase Ran/Gsp1p and Ntf2p is required for nuclear transport. Mol. Cell. Biol 17, 3755–3767
    OpenUrlAbstract/FREE Full Text
    1. Yang, J.,
    2. Bardes, E. S. G.,
    3. Moore, J. D.,
    4. Brennan, J.,
    5. Powers, M. A. and
    6. Kornbluth, S.
    (1998). Control of Cyclin B1 localization through regulated binding of the nuclear export factor CRM1. Genes Dev 12, 2131–2143
    OpenUrlAbstract/FREE Full Text
Back to top

 Download PDF

Email
Journal Article
The Saccharomyces cerevisiae cyclin Clb2p is targeted to multiple subcellular locations by cis- and trans-acting determinants
J.K. Hood, W.W. Hwang, P.A. Silver
Journal of Cell Science 2001 114: 589-597;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Introducing FocalPlane’s new Community Manager, Esperanza Agullo-Pascual

We are pleased to welcome Esperanza to the Journal of Cell Science team. The new Community Manager for FocalPlane, Esperanza is joining us from the Microscopy Core at Mount Sinai School of Medicine. Find out more about Esperanza in her introductory post over on FocalPlane.

New funding scheme supports sustainable events

As part of our Sustainable Conferencing Initiative, we are pleased to announce funding for organisers that seek to reduce the environmental footprint of their event. The next deadline to apply for a Scientific Meeting grant is 26 March 2021.

Read & Publish participation continues to grow

"Alongside pre-printing for early documentation of work, such mechanisms are particularly helpful for early-career researchers like me.”

Dr Chris MacDonald (University of York) shares his experience of publishing Open Access as part of our growing Read & Publish initiative. We now have over 150 institutions in 15 countries and four library consortia taking part – find out more and view our full list of participating institutions.

Cell scientist to watch: Romain Levayer

In an interview, Romain Levayer talks about starting his own lab, his love for preprints and his experience of balancing parenting with his research goals.

Live lactating mammary tissue

In a stunning video, Stewart et al. demonstrate warping of the alveolar unit due to basal cell-generated force as part of their recent work investigating roles for mechanically activated ion channels in lactation and involution.

Visit our YouTube channel to watch more videos from JCS, our sister journals and the Company.

JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.