Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 633K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Mountain, V. Articles by Compton, D. A. Search for Related Content PubMed PubMed Citation Articles by Mountain, V. Articles by Compton, D. A. Related Collections Related Article Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/1999//351 $5.00
The Journal of Cell Biology, Volume 147, Number 2, , 1999 351-366

The Kinesin-Related Protein, Hset, Opposes the Activity of Eg5 and Cross-Links Microtubules in the Mammalian Mitotic Spindle

^a Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755
^b Departments of Cell-Developmental Biology, Obstetrics-Gynecology, and Oregon Regional Primate Research Center, Oregon Health Sciences University, Beaverton, Oregon 97006
^c Rippel Electron Microscope Facility, Dartmouth College, Hanover, New Hampshire 03755
^d Department of Genetic Information, Division of Molecular Life Science, University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193, Japan
Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755.(603) 650-1128(603) 650-1990

duane.a.compton{at}dartmouth.edu

We have prepared antibodies specific for HSET, the human homologue of the KAR3 family of minus end-directed motors. Immuno-EM with these antibodies indicates that HSET frequently localizes between microtubules within the mammalian metaphase spindle consistent with a microtubule cross-linking function. Microinjection experiments show that HSET activity is essential for meiotic spindle organization in murine oocytes and taxol-induced aster assembly in cultured cells. However, inhibition of HSET did not affect mitotic spindle architecture or function in cultured cells, indicating that centrosomes mask the role of HSET during mitosis. We also show that (acentrosomal) microtubule asters fail to assemble in vitro without HSET activity, but simultaneous inhibition of HSET and Eg5, a plus end-directed motor, redresses the balance of forces acting on microtubules and restores aster organization. In vivo, centrosomes fail to separate and monopolar spindles assemble without Eg5 activity. Simultaneous inhibition of HSET and Eg5 restores centrosome separation and, in some cases, bipolar spindle formation. Thus, through microtubule cross-linking and oppositely oriented motor activity, HSET and Eg5 participate in spindle assembly and promote spindle bipolarity, although the activity of HSET is not essential for spindle assembly and function in cultured cells because of centrosomes.

Key Words: mitotic spindle • HSET • Eg5 • kinesin • microtubule

© 1999 The Rockefeller University Press

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• FitzHarris, G. (2009). A shift from kinesin 5-dependent metaphase spindle function during preimplantation development in mouse. Development 136: 2111-2119 [Abstract] [Full Text]  
• Moutinho-Pereira, S., Debec, A., Maiato, H. (2009). Microtubule Cytoskeleton Remodeling by Acentriolar Microtubule-organizing Centers at the Entry and Exit from Mitosis in Drosophila Somatic Cells. Mol. Biol. Cell 20: 2796-2808 [Abstract] [Full Text]  
• Cai, S., Weaver, L. N., Ems-McClung, S. C., Walczak, C. E. (2009). Kinesin-14 Family Proteins HSET/XCTK2 Control Spindle Length by Cross-Linking and Sliding Microtubules. Mol. Biol. Cell 20: 1348-1359 [Abstract] [Full Text]  
• Kwon, M., Godinho, S. A., Chandhok, N. S., Ganem, N. J., Azioune, A., Thery, M., Pellman, D. (2008). Mechanisms to suppress multipolar divisions in cancer cells with extra centrosomes. Genes Dev. 22: 2189-2203 [Abstract] [Full Text]  
• Krzysiak, T. C., Grabe, M., Gilbert, S. P. (2008). Getting in Sync with Dimeric Eg5: INITIATION AND REGULATION OF THE PROCESSIVE RUN. J. Biol. Chem. 283: 2078-2087 [Abstract] [Full Text]  
• Castillo, A., Morse, H. C. III, Godfrey, V. L., Naeem, R., Justice, M. J. (2007). Overexpression of Eg5 Causes Genomic Instability and Tumor Formation in Mice. Cancer Res. 67: 10138-10147 [Abstract] [Full Text]  
• Zhang, J., Megraw, T. L. (2007). Proper Recruitment of {gamma}-Tubulin and D-TACC/Msps to Embryonic Drosophila Centrosomes Requires Centrosomin Motif 1. Mol. Biol. Cell 18: 4037-4049 [Abstract] [Full Text]  
• Myers, K. A., Baas, P. W. (2007). Kinesin-5 regulates the growth of the axon by acting as a brake on its microtubule array. JCB 178: 1081-1091 [Abstract] [Full Text]  
• Oladipo, A., Cowan, A., Rodionov, V. (2007). Microtubule Motor Ncd Induces Sliding of Microtubules In Vivo. Mol. Biol. Cell 18: 3601-3606 [Abstract] [Full Text]  
• Bannigan, A., Scheible, W.-R., Lukowitz, W., Fagerstrom, C., Wadsworth, P., Somerville, C., Baskin, T. I. (2007). A conserved role for kinesin-5 in plant mitosis. J. Cell Sci. 120: 2819-2827 [Abstract] [Full Text]  
• Manning, A. L., Ganem, N. J., Bakhoum, S. F., Wagenbach, M., Wordeman, L., Compton, D. A. (2007). The Kinesin-13 Proteins Kif2a, Kif2b, and Kif2c/MCAK Have Distinct Roles during Mitosis in Human Cells. Mol. Biol. Cell 18: 2970-2979 [Abstract] [Full Text]  
• Barrett, S. L., Albertini, D. F. (2007). Allocation of Gamma-Tubulin Between Oocyte Cortex and Meiotic Spindle Influences Asymmetric Cytokinesis in the Mouse Oocyte. Biol. Reprod. 76: 949-957 [Abstract] [Full Text]  
• Hall, V.J., Compton, D., Stojkovic, P., Nesbitt, M., Herbert, M., Murdoch, A., Stojkovic, M. (2007). Developmental competence of human in vitro aged oocytes as host cells for nuclear transfer. Hum Reprod 22: 52-62 [Abstract] [Full Text]  
• Ambrose, J. C., Cyr, R. (2007). The Kinesin ATK5 Functions in Early Spindle Assembly in Arabidopsis. Plant Cell 19: 226-236 [Abstract] [Full Text]  
• McNally, K., Audhya, A., Oegema, K., McNally, F. J. (2006). Katanin controls mitotic and meiotic spindle length. JCB 175: 881-891 [Abstract] [Full Text]  
• Skoufias, D. A., DeBonis, S., Saoudi, Y., Lebeau, L., Crevel, I., Cross, R., Wade, R. H., Hackney, D., Kozielski, F. (2006). S-Trityl-L-cysteine Is a Reversible, Tight Binding Inhibitor of the Human Kinesin Eg5 That Specifically Blocks Mitotic Progression. J. Biol. Chem. 281: 17559-17569 [Abstract] [Full Text]  
• Christodoulou, A., Lederer, C. W., Surrey, T., Vernos, I., Santama, N. (2006). Motor protein KIFC5A interacts with Nubp1 and Nubp2, and is implicated in the regulation of centrosome duplication. J. Cell Sci. 119: 2035-2047 [Abstract] [Full Text]  
• Silverman-Gavrila, R. V., Wilde, A. (2006). Ran Is Required before Metaphase for Spindle Assembly and Chromosome Alignment and after Metaphase for Chromosome Segregation and Spindle Midbody Organization. Mol. Biol. Cell 17: 2069-2080 [Abstract] [Full Text]  
• Goshima, G., Nedelec, F., Vale, R. D. (2005). Mechanisms for focusing mitotic spindle poles by minus end-directed motor proteins. JCB 171: 229-240 [Abstract] [Full Text]  
• Zhu, C., Zhao, J., Bibikova, M., Leverson, J. D., Bossy-Wetzel, E., Fan, J.-B., Abraham, R. T., Jiang, W. (2005). Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference. Mol. Biol. Cell 16: 3187-3199 [Abstract] [Full Text]  
• Cochran, J. C., Gatial, J. E. III, Kapoor, T. M., Gilbert, S. P. (2005). Monastrol Inhibition of the Mitotic Kinesin Eg5. J. Biol. Chem. 280: 12658-12667 [Abstract] [Full Text]  
• Ambrose, J. C., Li, W., Marcus, A., Ma, H., Cyr, R. (2005). A Minus-End-directed Kinesin with Plus-End Tracking Protein Activity Is Involved in Spindle Morphogenesis. Mol. Biol. Cell 16: 1584-1592 [Abstract] [Full Text]  
• Quintyne, N. J., Reing, J. E., Hoffelder, D. R., Gollin, S. M., Saunders, W. S. (2005). Spindle Multipolarity Is Prevented by Centrosomal Clustering. Science 307: 127-129 [Abstract] [Full Text]  
• Manandhar, G., Schatten, H., Sutovsky, P. (2005). Centrosome Reduction During Gametogenesis and Its Significance. Biol. Reprod. 72: 2-13 [Abstract] [Full Text]  
• Wilson, P. G., Simmons, R., Shigali, S. (2004). Novel nuclear defects in KLP61F-deficient mutants in Drosophila are partially suppressed by loss of Ncd function. J. Cell Sci. 117: 4921-4933 [Abstract] [Full Text]  
• Cochran, J. C., Sontag, C. A., Maliga, Z., Kapoor, T. M., Correia, J. J., Gilbert, S. P. (2004). Mechanistic Analysis of the Mitotic Kinesin Eg5. J. Biol. Chem. 279: 38861-38870 [Abstract] [Full Text]  
• DeBonis, S., Skoufias, D. A., Lebeau, L., Lopez, R., Robin, G., Margolis, R. L., Wade, R. H., Kozielski, F. (2004). In vitro screening for inhibitors of the human mitotic kinesin Eg5 with antimitotic and antitumor activities. Molecular Cancer Therapeutics 3: 1079-1090 [Abstract] [Full Text]  
• Ganem, N. J., Compton, D. A. (2004). The KinI kinesin Kif2a is required for bipolar spindle assembly through a functional relationship with MCAK. JCB 166: 473-478 [Abstract] [Full Text]  
• Huang, S.-C., Jagadeeswaran, R., Liu, E. S., Benz, E. J. Jr. (2004). Protein 4.1R, a Microtubule-associated Protein Involved in Microtubule Aster Assembly in Mammalian Mitotic Extract. J. Biol. Chem. 279: 34595-34602 [Abstract] [Full Text]  
• Sakowicz, R., Finer, J. T., Beraud, C., Crompton, A., Lewis, E., Fritsch, A., Lee, Y., Mak, J., Moody, R., Turincio, R., Chabala, J. C., Gonzales, P., Roth, S., Weitman, S., Wood, K. W. (2004). Antitumor Activity of a Kinesin Inhibitor. Cancer Res. 64: 3276-3280 [Abstract] [Full Text]  
• Chakravarty, A., Howard, L., Compton, D. A. (2004). A Mechanistic Model for the Organization of Microtubule Asters by Motor and Non-Motor Proteins in a Mammalian Mitotic Extract. Mol. Biol. Cell 15: 2116-2132 [Abstract] [Full Text]  
• Ems-McClung, S. C., Zheng, Y., Walczak, C. E. (2004). Importin {alpha}/{beta} and Ran-GTP Regulate XCTK2 Microtubule Binding through a Bipartite Nuclear Localization Signal. Mol. Biol. Cell 15: 46-57 [Abstract] [Full Text]  
• Segbert, C., Barkus, R., Powers, J., Strome, S., Saxton, W. M., Bossinger, O. (2003). KLP-18, a Klp2 Kinesin, Is Required for Assembly of Acentrosomal Meiotic Spindles in Caenorhabditis elegans. Mol. Biol. Cell 14: 4458-4469 [Abstract] [Full Text]  
• Levesque, A. A., Howard, L., Gordon, M. B., Compton, D. A. (2003). A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells. Mol. Biol. Cell 14: 3541-3552 [Abstract] [Full Text]  
• Simerly, C., Dominko, T., Navara, C., Payne, C., Capuano, S., Gosman, G., Chong, K.-Y., Takahashi, D., Chace, C., Compton, D., Hewitson, L., Schatten, G. (2003). Molecular Correlates of Primate Nuclear Transfer Failures. Science 300: 297-297 [Full Text]  
• Khodjakov, A., Copenagle, L., Gordon, M. B., Compton, D. A., Kapoor, T. M. (2003). Minus-end capture of preformed kinetochore fibers contributes to spindle morphogenesis. JCB 160: 671-683 [Abstract] [Full Text]  
• Rusan, N. M., Tulu, U. S., Fagerstrom, C., Wadsworth, P. (2002). Reorganization of the microtubule array in prophase/prometaphase requires cytoplasmic dynein-dependent microtubule transport. JCB 158: 997-1003 [Abstract] [Full Text]  
• Roig, J., Mikhailov, A., Belham, C., Avruch, J. (2002). Nercc1, a mammalian NIMA-family kinase, binds the Ran GTPase and regulates mitotic progression. Genes Dev. 16: 1640-1658 [Abstract] [Full Text]  
• Haren, L., Merdes, A. (2002). Direct binding of NuMA to tubulin is mediated by a novel sequence motif in the tail domain that bundles and stabilizes microtubules. J. Cell Sci. 115: 1815-1824 [Abstract] [Full Text]  
• Troxell, C. L., Sweezy, M. A., West, R. R., Reed, K. D., Carson, B. D., Pidoux, A. L., Cande, W. Z., McIntosh, J. R. (2001). pkl1+and klp2+: Two Kinesins of the Kar3 Subfamily in Fission Yeast Perform Different Functions in Both Mitosis and Meiosis. Mol. Biol. Cell 12: 3476-3488 [Abstract] [Full Text]  
• Kapoor, T. M., Mitchison, T. J. (2001). Eg5 is static in bipolar spindles relative to tubulin: evidence for a static spindle matrix. JCB 154: 1125-1134 [Abstract] [Full Text]  
• Roy, L., Coullin, P., Vitrat, N., Hellio, R., Debili, N., Weinstein, J., Bernheim, A., Vainchenker, W. (2001). Asymmetrical segregation of chromosomes with a normal metaphase/anaphase checkpoint in polyploid megakaryocytes. Blood 97: 2238-2247 [Abstract] [Full Text]  
• Hu, Y., Betzendahl, I., Cortvrindt, R., Smitz, J., Eichenlaub-Ritter, U. (2001). Effects of low O2 and ageing on spindles and chromosomes in mouse oocytes from pre-antral follicle culture. Hum Reprod 16: 737-748 [Abstract] [Full Text]  
• Gordon, M. B., Howard, L., Compton, D. A. (2001). Chromosome Movement in Mitosis Requires Microtubule Anchorage at Spindle Poles. JCB 152: 425-434 [Abstract] [Full Text]  
• Touitou, I, Lhomond, G, Pruliere, G (2001). Boursin, a sea urchin bimC kinesin protein, plays a role in anaphase and cytokinesis. J. Cell Sci. 114: 481-491 [Abstract]  
• Kapoor, T. M., Mayer, T. U., Coughlin, M. L., Mitchison, T. J. (2000). Probing Spindle Assembly Mechanisms with Monastrol, a Small Molecule Inhibitor of the Mitotic Kinesin, Eg5. JCB 150: 975-988 [Abstract] [Full Text]  
• Rogers, G. C., Chui, K. K., Lee, E. W., Wedaman, K. P., Sharp, D. J., Holland, G., Morris, R. L., Scholey, J. M. (2000). A Kinesin-Related Protein, Krp180, Positions Prometaphase Spindle Poles during Early Sea Urchin Embryonic Cell Division. JCB 150: 499-512 [Abstract] [Full Text]  
• Dionne, M. A., Sanchez, A., Compton, D. A. (2000). ch-TOGp Is Required for Microtubule Aster Formation in a Mammalian Mitotic Extract. J. Biol. Chem. 275: 12346-12352 [Abstract] [Full Text]  
• Yvon, A.-M. C., Gross, D. J., Wadsworth, P. (2001). Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells. Proc. Natl. Acad. Sci. USA 98: 8656-8661 [Abstract] [Full Text]  
• Mack, G. J., Compton, D. A. (2001). Analysis of mitotic microtubule-associated proteins using mass spectrometry identifies astrin, a spindle-associated protein. Proc. Natl. Acad. Sci. USA 98: 14434-14439 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents