The Library

Latrunculin A delays anaphase onset in fission yeast by disrupting an ase1-independent pathway controlling mitotic spindle stability

Tools

Meadows, John C. and Millar, Jonathan B. A.. (2008) Latrunculin A delays anaphase onset in fission yeast by disrupting an ase1-independent pathway controlling mitotic spindle stability. Molecular Biology of the Cell, Vol.19 (No.9). pp. 3713-3723. ISSN 1059-1524

Preview

PDF
WRAP_Meadows_Latrunculin.pdf - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Download (1596Kb)

Official URL: http://dx.doi.org/10.1091/mbc.E08-02-0164

Abstract

It has been proposed previously that latrunculin A, an inhibitor of actin polymerization, delays the onset of anaphase by causing spindle misorientation in fission yeast. However, we show that {Delta}mto1 cells, which are defective in nucleation of cytoplasmic microtubules, have profoundly misoriented spindles but are not delayed in the timing of sister chromatid separation, providing compelling evidence that fission yeast does not possess a spindle orientation checkpoint. Instead, we show that latrunculin A delays anaphase onset by disrupting interpolar microtubule stability. This effect is abolished in a latrunculin A-insensitive actin mutant and exacerbated in cells lacking Ase1, which cross-links antiparallel interpolar microtubules at the spindle midzone both before and after anaphase. These data indicate that both Ase1 and an intact actin cytoskeleton are required for preanaphase spindle stability. Finally, we show that loss of Ase1 activates a checkpoint that requires only the Mad3, Bub1, and Mph1, but not Mad1, Mad2, or Bub3 checkpoint proteins.

Item Type:	Journal Article
Subjects:	Q Science > QK Botany Q Science > QR Microbiology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Medicine > Warwick Medical School > Biomedical Cell Biology Faculty of Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	Actin, Polymerization, Nucleation, Microtubules, Mitosis -- Regulation
Journal or Publication Title:	Molecular Biology of the Cell
Publisher:	American Society for Cell Biology
ISSN:	1059-1524
Date:	September 2008
Volume:	Vol.19
Number:	No.9
Number of Pages:	11
Page Range:	pp. 3713-3723
Identification Number:	10.1091/mbc.E08-02-0164
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Association for International Cancer Research (AICR), Medical Research Council (Great Britain) (MRC)
References:	Asakawa, K., Kume, K., Kanai, M., Goshima, T., Miyahara, K., Dhut, S., Tee, W. W., Hirata, D., and Toda, T. (2006). The V260I mutation in fission yeast alpha-tubulin Atb2 affects microtubule dynamics and EB1-Mal3 localization and activates the Bub1 branch of the spindle checkpoint. Mol. Biol. Cell 17, 1421–1435. Asakawa, K., Toya, M., Sato, M., Kanai, M., Kume, K., Goshima, T., Garcia, M. A., Hirata, D., and Toda, T. (2005). Mal3, the fission yeast EB1 homologue, cooperates with Bub1 spindle checkpoint to prevent monopolar attachment. EMBO Rep 6, 1194–1200. Ayscough, K. R., Stryker, J., Pokala, N., Sanders, M., Crews, P., and Drubin, D. G. (1997). High rates of actin filament turnover in budding yeast and roles for actin in establishment and maintenance of cell polarity revealed using the actin inhibitor latrunculin-A. J. Cell Biol 137, 399–416. Bahler, J., Wu, J. Q., Longtine, M. S., Shah, N. G., McKenzie, A., 3rd, Steever, A. B., Wach, A., Philippsen, P., and Pringle, J. R. (1998). Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast 14, 943–951. Bernard, P., Hardwick, K., and Javerzat, J. P. (1998). Fission yeast bub1 is a mitotic centromere protein essential for the spindle checkpoint and the preservation of correct ploidy through mitosis. J. Cell Biol 143, 1775–1787. Bratman, S. V., and Chang, F. (2007). Stabilization of overlapping microtubules by fission yeast CLASP. Dev. Cell 13, 812–827. Chan, J., Jensen, C. G., Jensen, L. C., Bush, M., and Lloyd, C. W. (1999). The 65-kDa carrot microtubule-associated protein forms regularly arranged filamentous cross-bridges between microtubules. Proc. Natl. Acad. Sci. USA 96, 14931–14936. Chen, R. H., Waters, J. C., Salmon, E. D., and Murray, A. W. (1996). Association of spindle assembly checkpoint component XMAD2 with unattached kinetochores. Science 274, 242–246. Cleveland, D. W., Mao, Y., and Sullivan, K. F. (2003). Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling. Cell 112, 407–421. Ding, R., McDonald, K. L., and McIntosh, J. R. (1993). Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. J. Cell Biol 120, 141–151. Fabian, L., Xia, X., Venkitaramani, D. V., Johansen, K. M., Johansen, J., Andrew, D. J., and Forer, A. (2007). Titin in insect spermatocyte spindle fibers associates with microtubules, actin, myosin and the matrix proteins skeletor, megator and chromator. J. Cell Sci 120, 2190–2204. Fang, G. (2002). Checkpoint protein BubR1 acts synergistically with Mad2 to inhibit anaphase-promoting complex. Mol. Biol. Cell 13, 755–766. Fujita, M., Ichinose, S., Kiyono, T., Tsurumi, T., and Omori, A. (2003). Establishment of latrunculin-A resistance in HeLa cells by expression of R183A D184A mutant beta-actin. Oncogene 22, 627–631. Funabiki, H., Hagan, I., Uzawa, S., and Yanagida, M. (1993). Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast. J. Cell Biol 121, 961–976. Gachet, Y., Tournier, S., Millar, J. B., and Hyams, J. S. (2001). A MAP kinase-dependent actin checkpoint ensures proper spindle orientation in fission yeast. Nature 412, 352–355. Gachet, Y., Tournier, S., Millar, J. B., and Hyams, J. S. (2004). Mechanism controlling perpendicular alignment of the spindle to the axis of cell division in fission yeast. EMBO J 23, 1289–1300. Gillett, E. S., Espelin, C. W., and Sorger, P. K. (2004). Spindle checkpoint proteins and chromosome-microtubule attachment in budding yeast. J. Cell Biol 164, 535–546. Grallert, A., Beuter, C., Craven, R. A., Bagley, S., Wilks, D., Fleig, U., and Hagan, I. M. (2006). S. pombe CLASP needs dynein, not EB1 or CLIP170, to induce microtubule instability and slows polymerization rates at cell tips in a dynein-dependent manner. Genes Dev 20, 2421–2436. Hagan, I. M. (1998). The fission yeast microtubule cytoskeleton. J. Cell Sci 111, 1603–1612. Hagan, I. M., and Hyams, J. S. (1996). Forces acting on the fission yeast anaphase spindle. Cell Motil. Cytoskeleton 34, 69–75. He, X., Jones, M. H., Winey, M., and Sazer, S. (1998). Mph1, a member of the Mps1-like family of dual specificity protein kinases, is required for the spindle checkpoint in S. pombe. J. Cell Sci 111, 1635–1647. He, X., Patterson, T. E., and Sazer, S. (1997). The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex. Proc. Natl. Acad. Sci. USA 94, 7965–7970. Hoyt, M. A., Totis, L., and Roberts, B. T. (1991). S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 66, 507–517. Hwang, L. H., Lau, L. F., Smith, D. L., Mistrot, C. A., Hardwick, K. G., Hwang, E. S., Amon, A., and Murray, A. W. (1998). Budding yeast Cdc 20, a target of the spindle checkpoint. Science 279, 1041–1044. Jiang, W., Jimenez, G., Wells, N. J., Hope, T. J., Wahl, G. M., Hunter, T., and Fukunaga, R. (1998). PRC 1, a human mitotic spindle-associated CDK substrate protein required for cytokinesis. Mol. Cell 2, 877–885. Khmelinskii, A., Lawrence, C., Roostalu, J., and Schiebel, E. (2007). Cdc14-regulated midzone assembly controls anaphase B. J. Cell Biol 177, 981–993. Kim, S. H., Lin, D. P., Matsumoto, S., Kitazono, A., and Matsumoto, T. (1998). Fission yeast Slp 1, an effector of the Mad2-dependent spindle checkpoint. Science 279, 1045–1047. King, E. M., Rachidi, N., Morrice, N., Hardwick, K. G., and Stark, M. J. (2007). Ipl1p-dependent phosphorylation of Mad3p is required for the spindle checkpoint response to lack of tension at kinetochores. Genes Dev 21, 1163–1168. Kotwaliwale, C. V., Frei, S. B., Stern, B. M., and Biggins, S. (2007). A pathway containing the Ipl1/aurora protein kinase and the spindle midzone protein Ase1 regulates yeast spindle assembly. Dev. Cell 13, 433–445. Krapp, A., Schmidt, S., Cano, E., and Simanis, V. (2001). S. pombe cdc11p, together with sid4p, provides an anchor for septation initiation network proteins on the spindle pole body. Curr. Biol 11, 1559–1568. Lenart, P., Bacher, C. P., Daigle, N., Hand, A. R., Eils, R., Terasaki, M., and Ellenberg, J. (2005). A contractile nuclear actin network drives chromosome congression in oocytes. Nature 436, 812–818. Li, R., and Murray, A. W. (1991). Feedback control of mitosis in budding yeast. Cell 66, 519–531. Li, X., and Nicklas, R. B. (1995). Mitotic forces control a cell-cycle checkpoint. Nature 373, 630–632. Loiodice, I., Staub, J., Setty, T. G., Nguyen, N. P., Paoletti, A., and Tran, P. T. (2005). Ase1p organizes antiparallel microtubule arrays during interphase and mitosis in fission yeast. Mol. Biol. Cell 16, 1756–1768. Marks, J., Hagan, I. M., and Hyams, J. S. (1986). Growth polarity and cytokinesis in fission yeast: the role of the cytoskeleton. J. Cell Sci. Suppl 5, 229–241. Millband, D. N., and Hardwick, K. G. (2002). Fission yeast Mad3p is required for Mad2p to inhibit the anaphase-promoting complex and localizes to kinetochores in a Bub1p-, Bub3p-, and Mph1p-dependent manner. Mol. Cell Biol 22, 2728–2742. Nabeshima, K., Nakagawa, T., Straight, A. F., Murray, A., Chikashige, Y., Yamashita, Y. M., Hiraoka, Y., and Yanagida, M. (1998). Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis1 is implicated in force balance in metaphase bipolar spindle. Mol. Biol. Cell 9, 3211–3225. Oliferenko, S., and Balasubramanian, M. K. (2002). Astral microtubules monitor metaphase spindle alignment in fission yeast. Nat. Cell Biol 4, 816–820. Orr, B., Bousbaa, H., and Sunkel, C. E. (2007). Mad2-independent spindle assembly checkpoint activation and controlled metaphase-anaphase transition in Drosophila S2 cells. Mol. Biol. Cell 18, 850–863. Rajagopalan, S., Bimbo, A., Balasubramanian, M. K., and Oliferenko, S. (2004). A potential tension-sensing mechanism that ensures timely anaphase onset upon metaphase spindle orientation. Curr. Biol 14, 69–74. Rieder, C. L., Cole, R. W., Khodjakov, A., and Sluder, G. (1995). The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J. Cell Biol 130, 941–948. Rupes, I., Webb, B. A., Mak, A., and Young, P. G. (2001). G2/M arrest caused by actin disruption is a manifestation of the cell size checkpoint in fission yeast. Mol. Biol. Cell 12, 3892–3903. Sato, M., Koonrugsa, N., Toda, T., Vardy, L., Tournier, S., and Millar, J. B. (2003). Deletion of Mia1/Alp7 activates Mad2-dependent spindle assembly checkpoint in fission yeast. Nat. Cell Biol 5, 764–766. author reply 766. Sawin, K. E., Lourenco, P. C., and Snaith, H. A. (2004). Microtubule nucleation at non-spindle pole body microtubule-organizing centers requires fission yeast centrosomin-related protein mod20p. Curr. Biol 14, 763–775. Sawin, K. E., and Nurse, P. (1998). Regulation of cell polarity by microtubules in fission yeast. J. Cell Biol 142, 457–471. Schuyler, S. C., Liu, J. Y., and Pellman, D. (2003). The molecular function of Ase1p: evidence for a MAP-dependent midzone-specific spindle matrix. Microtubule-associated proteins. J. Cell Biol 160, 517–528. Silverman-Gavrila, R. V., and Forer, A. (2000). Evidence that actin and myosin are involved in the poleward flux of tubulin in metaphase kinetochore microtubules of crane-fly spermatocytes. J. Cell Sci 113, 597–609. Skoufias, D. A., Andreassen, P. R., Lacroix, F. B., Wilson, L., and Margolis, R. L. (2001). Mammalian mad2 and bub1/bubR1 recognize distinct spindle-attachment and kinetochore-tension checkpoints. Proc. Natl. Acad. Sci. USA 98, 4492–4497. Stern, B. M., and Murray, A. W. (2001). Lack of tension at kinetochores activates the spindle checkpoint in budding yeast. Curr. Biol 11, 1462–1467. Sudakin, V., Chan, G. K., and Yen, T. J. (2001). Checkpoint inhibition of the APC/C in HeLa cells is mediated by a complex of BUBR1, BUB3, CDC20, and MAD2. J. Cell Biol 154, 925–936. Tang, Z., Bharadwaj, R., Li, B., and Yu, H. (2001). Mad2-Independent inhibition of APCCdc20 by the mitotic checkpoint protein BubR1. Dev. Cell 1, 227–237. Tange, Y., and Niwa, O. (2008). Schizosaccharomyces pombe Bub3 is dispensable for mitotic arrest following perturbed spindle formation. Genetics 179, 785–792. Tatebe, H., Goshima, G., Takeda, K., Nakagawa, T., Kinoshita, K., and Yanagida, M. (2001). Fission yeast living mitosis visualized by GFP-tagged gene products. Micron 32, 67–74. Tournier, S., Gachet, Y., Buck, V., Hyams, J. S., and Millar, J. B. (2004). Disruption of astral microtubule contact with the cell cortex activates a Bub1, Bub3, and Mad3-dependent checkpoint in fission yeast. Mol. Biol. Cell 15, 3345–3356. Toya, M., Sato, M., Haselmann, U., Asakawa, K., Brunner, D., Antony, C., and Toda, T. (2007). Gamma-tubulin complex-mediated anchoring of spindle microtubules to spindle-pole bodies requires Msd1 in fission yeast. Nat. Cell Biol 9, 646–653. Tran, P. T., Marsh, L., Doye, V., Inoue, S., and Chang, F. (2001). A mechanism for nuclear positioning in fission yeast based on microtubule pushing. J. Cell Biol 153, 397–411. Uhlmann, F. (2004). The mechanism of sister chromatid cohesion. Exp. Cell Res 296, 80–85. van Hemert, M. J., Lamers, G. E., Klein, D. C., Oosterkamp, T. H., Steensma, H. Y., and van Heusden, G. P. (2002). The Saccharomyces cerevisiae Fin1 protein forms cell cycle-specific filaments between spindle pole bodies. Proc. Natl. Acad. Sci. USA 99, 5390–5393. Vanoosthuyse, V., Valsdottir, R., Javerzat, J. P., and Hardwick, K. G. (2004). Kinetochore targeting of fission yeast Mad and Bub proteins is essential for spindle checkpoint function but not for all chromosome segregation roles of Bub1p. Mol. Cell Biol 24, 9786–9801. Venkatram, S., Tasto, J. J., Feoktistova, A., Jennings, J. L., Link, A. J., and Gould, K. L. (2004). Identification and characterization of two novel proteins affecting fission yeast gamma-tubulin complex function. Mol. Biol. Cell 15, 2287–2301. Vogel, S. K., Raabe, I., Dereli, A., Maghelli, N., and Tolic-Norrelykke, I. (2007). Interphase microtubules determine the initial alignment of the mitotic spindle. Curr. Biol 17, 438–444. Waters, J. C., Chen, R. H., Murray, A. W., and Salmon, E. D. (1998). Localization of Mad2 to kinetochores depends on microtubule attachment, not tension. J. Cell Biol 141, 1181–1191. Weiss, E., and Winey, M. (1996). The Saccharomyces cerevisiae spindle pole body duplication gene MPS1 is part of a mitotic checkpoint. J. Cell Biol 132, 111–123. Wigge, P. A., and Kilmartin, J. V. (2001). The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation. J. Cell Biol 152, 349–360. Woodbury, E. L., and Morgan, D. O. (2007). Cdk and APC activities limit the spindle-stabilizing function of Fin1 to anaphase. Nat. Cell Biol 9, 106–112. Yamashita, A., Sato, M., Fujita, A., Yamamoto, M., and Toda, T. (2005). The roles of fission yeast ase1 in mitotic cell division, meiotic nuclear oscillation, and cytokinesis checkpoint signaling. Mol. Biol. Cell 16, 1378–1395. Zhou, J., Yao, J., and Joshi, H. C. (2002). Attachment and tension in the spindle assembly checkpoint. J. Cell Sci 115, 3547–3555. Zhu, C., Lau, E., Schwarzenbacher, R., Bossy-Wetzel, E., and Jiang, W. (2006). Spatiotemporal control of spindle midzone formation by PRC1 in human cells. Proc. Natl. Acad. Sci. USA 103, 6196–6201. Zimmerman, S., and Chang, F. (2005). Effects of {gamma}-tubulin complex proteins on microtubule nucleation and catastrophe in fission yeast. Mol. Biol. Cell 16, 2719–2733. Zimmerman, S., Daga, R. R., and Chang, F. (2004). Intra-nuclear microtubules and a mitotic spindle orientation checkpoint. Nat. Cell Biol 6, 1245–1246.
URI:	http://wrap.warwick.ac.uk/id/eprint/942