J. Cell Biol., Volume 141, Number 3, May 4, 1998 663-674

The Mammalian -Tubulin Complex Contains Homologues of the Yeast Spindle Pole Body Components Spc97p and Spc98p

Steven M. Murphy, Lenore Urbani, and Tim Stearns

Department of Biological Sciences, Stanford University, Stanford, California

-Tubulin is a universal component of microtubule organizing centers where it is believed to play an important role in the nucleation of microtubule polymerization. -Tubulin also exists as part of a cytoplasmic complex whose size and complexity varies in different organisms. To investigate the composition of the cytoplasmic -tubulin complex in mammalian cells, cell lines stably expressing epitope-tagged versions of human -tubulin were made. The epitope-tagged -tubulins expressed in these cells localize to the centrosome and are incorporated into the cytoplasmic -tubulin complex. Immunoprecipitation of this complex identifies at least seven proteins, with calculated molecular weights of 48, 71, 76, 100, 101, 128, and 211 kD. We have identified the 100- and 101-kD components of the -tubulin complex as homologues of the yeast spindle pole body proteins Spc97p and Spc98p, and named the corresponding human proteins hGCP2 and hGCP3. Sequence analysis revealed that these proteins are not only related to their respective homologues, but are also related to each other. GCP2 and GCP3 colocalize with -tubulin at the centrosome, cosediment with -tubulin in sucrose gradients, and coimmunoprecipitate with -tubulin, indicating that they are part of the -tubulin complex. The conservation of a complex involving -tubulin, GCP2, and GCP3 from yeast to mammals suggests that structurally diverse microtubule organizing centers such as the yeast spindle pole body and the animal centrosome share a common molecular mechanism for microtubule nucleation.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Izumi, N., Fumoto, K., Izumi, S., Kikuchi, A. (2008). GSK-3{beta} Regulates Proper Mitotic Spindle Formation in Cooperation with a Component of the {gamma}-Tubulin Ring Complex, GCP5. J. Biol. Chem. 283: 12981-12991 [Abstract] [Full Text]  
• Cunningham, L. A., Kahn, R. A. (2008). Cofactor D Functions as a Centrosomal Protein and Is Required for the Recruitment of the {gamma}-Tubulin Ring Complex at Centrosomes and Organization of the Mitotic Spindle. J. Biol. Chem. 283: 7155-7165 [Abstract] [Full Text]  
• Wiese, C. (2008). Distinct Dgrip84 Isoforms Correlate with Distinct {gamma}-Tubulins in Drosophila. Mol. Biol. Cell 19: 368-377 [Abstract] [Full Text]  
• Hoppeler-Lebel, A., Celati, C., Bellett, G., Mogensen, M. M., Klein-Hitpass, L., Bornens, M., Tassin, A.-M. (2007). Centrosomal CAP350 protein stabilises microtubules associated with the Golgi complex. J. Cell Sci. 120: 3299-3308 [Abstract] [Full Text]  
• Anders, A., Lourenco, P. C.C., Sawin, K. E. (2006). Noncore Components of the Fission Yeast {gamma}-Tubulin Complex. Mol. Biol. Cell 17: 5075-5093 [Abstract] [Full Text]  
• Vogt, N., Koch, I., Schwarz, H., Schnorrer, F., Nusslein-Volhard, C. (2006). The {gamma}TuRC components Grip75 and Grip128 have an essential microtubule-anchoring function in the Drosophila germline. Development 133: 3963-3972 [Abstract] [Full Text]  
• Sulimenko, V., Draberova, E., Sulimenko, T., Macurek, L., Richterova, V., Draber, P., Draber, P. (2006). Regulation of Microtubule Formation in Activated Mast Cells by Complexes of {gamma}-Tubulin with Fyn and Syk Kinases.. J. Immunol. 176: 7243-7253 [Abstract] [Full Text]  
• Wang, Q., Du, X., Meinkoth, J., Hirohashi, Y., Zhang, H., Liu, Q., Richter, M., Greene, M. I. (2006). Characterization of Su48, a centrosome protein essential for cell division. Proc. Natl. Acad. Sci. USA 103: 6512-6517 [Abstract] [Full Text]  
• Haren, L., Remy, M.-H., Bazin, I., Callebaut, I., Wright, M., Merdes, A. (2006). NEDD1-dependent recruitment of the {gamma}-tubulin ring complex to the centrosome is necessary for centriole duplication and spindle assembly. J. Cell Biol. 172: 505-515 [Abstract] [Full Text]  
• Verollet, C., Colombie, N., Daubon, T., Bourbon, H.-M., Wright, M., Raynaud-Messina, B. (2006). Drosophila melanogaster {gamma}-TuRC is dispensable for targeting {gamma}-tubulin to the centrosome and microtubule nucleation. J. Cell Biol. 172: 517-528 [Abstract] [Full Text]  
• Colombie, N., Verollet, C., Sampaio, P., Moisand, A., Sunkel, C., Bourbon, H.-M., Wright, M., Raynaud-Messina, B. (2006). The Drosophila {gamma}-Tubulin Small Complex Subunit Dgrip84 Is Required for Structural and Functional Integrity of the Spindle Apparatus. Mol. Biol. Cell 17: 272-282 [Abstract] [Full Text]  
• Delgehyr, N., Sillibourne, J., Bornens, M. (2005). Microtubule nucleation and anchoring at the centrosome are independent processes linked by ninein function. J. Cell Sci. 118: 1565-1575 [Abstract] [Full Text]  
• Barr, F. A., Egerer, J. (2005). Golgi positioning: are we looking at the right MAP?. J. Cell Biol. 168: 993-998 [Abstract] [Full Text]  
• Raynaud-Messina, B., Mazzolini, L., Moisand, A., Cirinesi, A.-M., Wright, M. (2004). Elongation of centriolar microtubule triplets contributes to the formation of the mitotic spindle in {gamma}-tubulin-depleted cells. J. Cell Sci. 117: 5497-5507 [Abstract] [Full Text]  
• Zimmerman, W. C., Sillibourne, J., Rosa, J., Doxsey, S. J. (2004). Mitosis-specific Anchoring of {gamma} Tubulin Complexes by Pericentrin Controls Spindle Organization and Mitotic Entry. Mol. Biol. Cell 15: 3642-3657 [Abstract] [Full Text]  
• Tange, Y., Fujita, A., Toda, T., Niwa, O. (2004). Functional Dissection of the {gamma}-Tubulin Complex by Suppressor Analysis of gtb1 and alp4 Mutations in Schizosaccharomyces pombe. Genetics 167: 1095-1107 [Abstract] [Full Text]  
• Venkatram, S., Tasto, J. J., Feoktistova, A., Jennings, J. L., Link, A. J., Gould, K. L. (2004). Identification and Characterization of Two Novel Proteins Affecting Fission Yeast {gamma}-tubulin Complex Function. Mol. Biol. Cell 15: 2287-2301 [Abstract] [Full Text]  
• You, Y., Huang, T., Richer, E. J., Schmidt, J.-E. H., Zabner, J., Borok, Z., Brody, S. L. (2004). Role of f-box factor foxj1 in differentiation of ciliated airway epithelial cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 286: L650-L657 [Abstract] [Full Text]  
• Kawaguchi, S.-i., Zheng, Y. (2004). Characterization of a Drosophila Centrosome Protein CP309 That Shares Homology with Kendrin and CG-NAP. Mol. Biol. Cell 15: 37-45 [Abstract] [Full Text]  
• Keryer, G., Di Fiore, B., Celati, C., Lechtreck, K. F., Mogensen, M., Delouvee, A., Lavia, P., Bornens, M., Tassin, A.-M. (2003). Part of Ran Is Associated with AKAP450 at the Centrosome: Involvement in Microtubule-organizing Activity. Mol. Biol. Cell 14: 4260-4271 [Abstract] [Full Text]  
• Gromley, A., Jurczyk, A., Sillibourne, J., Halilovic, E., Mogensen, M., Groisman, I., Blomberg, M., Doxsey, S. (2003). A novel human protein of the maternal centriole is required for the final stages of cytokinesis and entry into S phase. J. Cell Biol. 161: 535-545 [Abstract] [Full Text]  
• Gunawardane, R. N., Martin, O. C., Zheng, Y. (2003). Characterization of a New gamma TuRC Subunit with WD Repeats. Mol. Biol. Cell 14: 1017-1026 [Abstract] [Full Text]  
• Takahashi, M., Yamagiwa, A., Nishimura, T., Mukai, H., Ono, Y. (2002). Centrosomal Proteins CG-NAP and Kendrin Provide Microtubule Nucleation Sites by Anchoring gamma -Tubulin Ring Complex. Mol. Biol. Cell 13: 3235-3245 [Abstract] [Full Text]  
• Fujita, A., Vardy, L., Garcia, M. A., Toda, T. (2002). A Fourth Component of the Fission Yeast gamma -Tubulin Complex, Alp16, Is Required for Cytoplasmic Microtubule Integrity and Becomes Indispensable When gamma -Tubulin Function Is Compromised. Mol. Biol. Cell 13: 2360-2373 [Abstract] [Full Text]  
• Vinh, D. B.N., Kern, J. W., Hancock, W. O., Howard, J., Davis, T. N. (2002). Reconstitution and Characterization of Budding Yeast gamma -Tubulin Complex. Mol. Biol. Cell 13: 1144-1157 [Abstract] [Full Text]  
• Flory, M. R., Morphew, M., Joseph, J. D., Means, A. R., Davis, T. N. (2002). Pcp1p, an Spc110p-related Calmodulin Target at the Centrosome of the Fission Yeast Schizosaccharomyces pombe. Cell Growth Differ. 13: 47-58 [Abstract] [Full Text]  
• Erhardt, M., Stoppin-Mellet, V., Campagne, S., Canaday, J., Mutterer, J., Fabian, T., Sauter, M., Muller, T., Peter, C., Lambert, A.-M., Schmit, A.-C. (2002). The plant Spc98p homologue colocalizes with {gamma}-tubulin at microtubule nucleation sites and is required for microtubule nucleation. J. Cell Sci. 115: 2423-2431 [Abstract] [Full Text]  
• Hoffner, G., Kahlem, P., Djian, P. (2002). Perinuclear localization of huntingtin as a consequence of its binding to microtubules through an interaction with {beta}-tubulin: relevance to Huntington's disease. J. Cell Sci. 115: 941-948 [Abstract] [Full Text]  
• Murphy, S. M., Preble, A. M., Patel, U. K., O'Connell, K. L., Dias, D. P., Moritz, M., Agard, D., Stults, J. T., Stearns, T. (2001). GCP5 and GCP6: Two New Members of the Human gamma -Tubulin Complex. Mol. Biol. Cell 12: 3340-3352 [Abstract] [Full Text]  
• Nakagawa, Y., Yamane, Y., Okanoue, T., Tsukita, S., Tsukita, S. (2001). Outer Dense Fiber 2 Is a Widespread Centrosome Scaffold Component Preferentially Associated with Mother Centrioles: Its Identification from Isolated Centrosomes. Mol. Biol. Cell 12: 1687-1697 [Abstract] [Full Text]  
• Inclan, Y., Nogales, E (2001). Structural models for the self-assembly and microtubule interactions of gamma-, delta- and epsilon-tubulin. J. Cell Sci. 114: 413-422 [Abstract]  
• Gunawardane, R. N., Martin, O. C., Cao, K., Zhang, L., Dej, K., Iwamatsu, A., Zheng, Y. (2000). Characterization and Reconstitution of Drosophila {{gamma}}-Tubulin Ring Complex Subunits. J. Cell Biol. 151: 1513-1524 [Abstract] [Full Text]  
• Barbosa, V., Yamamoto, R. R., Henderson, D. S., Glover, D. M. (2000). Mutation of a Drosophila gamma tubulin ring complex subunit encoded by discs degenerate-4 differentially disrupts centrosomal protein localization. Genes Dev. 14: 3126-3139 [Abstract] [Full Text]  
• Takada, S., Shibata, T., Hiraoka, Y., Masuda, H. (2000). Identification of Ribonucleotide Reductase Protein R1 as an Activator of Microtubule Nucleation in Xenopus Egg Mitotic Extracts. Mol. Biol. Cell 11: 4173-4187 [Abstract] [Full Text]  
• Hung, L.-Y., Tang, C.-J. C., Tang, T. K. (2000). Protein 4.1 R-135 Interacts with a Novel Centrosomal Protein (CPAP) Which Is Associated with the gamma -Tubulin Complex. Mol. Cell. Biol. 20: 7813-7825 [Abstract] [Full Text]  
• Lin, H., Keriel, A., Morales, C. R., Bedard, N., Zhao, Q., Hingamp, P., Lefrançois, S., Combaret, L., Wing, S. S. (2000). Divergent N-Terminal Sequences Target an Inducible Testis Deubiquitinating Enzyme to Distinct Subcellular Structures. Mol. Cell. Biol. 20: 6568-6578 [Abstract] [Full Text]  
• Paluh, J. L., Nogales, E., Oakley, B. R., McDonald, K., Pidoux, A. L., Cande, W. Z. (2000). A Mutation in gamma -Tubulin Alters Microtubule Dynamics and Organization and Is Synthetically Lethal with the Kinesin-like Protein Pkl1p. Mol. Biol. Cell 11: 1225-1239 [Abstract] [Full Text]  
• Middendorp, S., Kuntziger, T., Abraham, Y., Holmes, S., Bordes, N., Paintrand, M., Paoletti, A., Bornens, M. (2000). A Role for Centrin 3 in Centrosome Reproduction. J. Cell Biol. 148: 405-416 [Abstract] [Full Text]  
• Schnackenberg, B., Hull, D., Balczon, R., Palazzo, R. (2000). Reconstitution of microtubule nucleation potential in centrosomes isolated from Spisula solidissima oocytes. J. Cell Sci. 113: 943-953 [Abstract]  
• Fava, F., Raynaud-Messina, B., Leung-Tack, J., Mazzolini, L., Li, M., Guillemot, J. C., Cachot, D., Tollon, Y., Ferrara, P., Wright, M. (1999). Human 76p: A New Member of the {gamma}-Tubulin-associated Protein Family. J. Cell Biol. 147: 857-868 [Abstract] [Full Text]  
• Draberova, L., Draberova, E., Surviladze, Z., Draber, P., Draber, P. (1999). Protein tyrosine kinase p53/p56lyn forms complexes with {gamma}-tubulin in rat basophilic leukemia cells. Int Immunol 11: 1829-1839 [Abstract] [Full Text]  
• Ashburner, M., Misra, S., Roote, J., Lewis, S. E., Blazej, R., Davis, T., Doyle, C., Galle, R., George, R., Harris, N., Hartzell, G., Harvey, D., Hong, L., Houston, K., Hoskins, R., Johnson, G., Martin, C., Moshrefi, A., Palazzolo, M., Reese, M. G., Spradling, A., Tsang, G., Wan, K., Whitelaw, K., Kimmel, B., Celniker, S., Rubin, G. M. (1999). An Exploration of the Sequence of a 2.9-Mb Region of the Genome of Drosophila melanogaster: The Adh Region. Genetics 153: 179-219 [Abstract] [Full Text]  
• Freed, E., Lacey, K. R., Huie, P., Lyapina, S. A., Deshaies, R. J., Stearns, T., Jackson, P. K. (1999). Components of an SCF ubiquitin ligase localize to the centrosome and regulate the centrosome duplication cycle. Genes Dev. 13: 2242-2257 [Abstract] [Full Text]  
• Adams, I. R., Kilmartin, J. V. (1999). Localization of Core Spindle Pole Body (SPB) Components during SPB Duplication in Saccharomyces cerevisiae. J. Cell Biol. 145: 809-823 [Abstract] [Full Text]  
• Oegema, K., Wiese, C., Martin, O. C., Milligan, R. A., Iwamatsu, A., Mitchison, T. J., Zheng, Y. (1999). Characterization of Two Related Drosophila gamma -tubulin Complexes that Differ in Their Ability to Nucleate Microtubules. J. Cell Biol. 144: 721-733 [Abstract] [Full Text]  
• Feierbach, B., Nogales, E., Downing, K. H., Stearns, T. (1999). Alf1p, a CLIP-170 Domain-containing Protein, Is Functionally and Physically Associated with alpha -Tubulin. J. Cell Biol. 144: 113-124 [Abstract] [Full Text]  
• Llamazares, S, Tavosanis, G, Gonzalez, C (1999). Cytological characterisation of the mutant phenotypes produced during early embryogenesis by null and loss-of-function alleles of the gammaTub37C gene in Drosophila. J. Cell Sci. 112: 659-667 [Abstract]  
• Bobinnec, Y., Khodjakov, A., Mir, L.M., Rieder, C.L., Edde, B., Bornens, M. (1998). Centriole Disassembly In Vivo and Its Effect on Centrosome Structure and Function in Vertebrate Cells. J. Cell Biol. 143: 1575-1589 [Abstract] [Full Text]  
• Qian, Y., Erikson, E., Maller, J. L. (1998). Purification and Cloning of a Protein Kinase That Phosphorylates and Activates the Polo-Like Kinase Plx1. Science 282: 1701-1704 [Abstract] [Full Text]  
• Nakamura, M., Masuda, H., Horii, J., Kuma, K.-i., Yokoyama, N., Ohba, T., Nishitani, H., Miyata, T., Tanaka, M., Nishimoto, T. (1998). When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to gamma -Tubulin. J. Cell Biol. 143: 1041-1052 [Abstract] [Full Text]  
• Hsu, L.-C., White, R. L. (1998). BRCA1 is associated with the centrosome during mitosis. Proc. Natl. Acad. Sci. USA 95: 12983-12988 [Abstract] [Full Text]  
• Leguy, R., Melki, R., Pantaloni, D., Carlier, M.-F. (2000). Monomeric gamma -Tubulin Nucleates Microtubules. J. Biol. Chem. 275: 21975-21980 [Abstract] [Full Text]  
• Hoepfner, D., Schaerer, F., Brachat, A., Wach, A., Philippsen, P. (2002). Reorientation of Mispositioned Spindles in Short Astral Microtubule Mutant spc72Delta Is Dependent on Spindle Pole Body Outer Plaque and Kar3 Motor Protein. Mol. Biol. Cell 13: 1366-1380 [Abstract] [Full Text]  

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