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

This Article Full Text Full Text (PDF, 2125K) 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 Vogel, J. M. Articles by Palazzo, R. E. Search for Related Content PubMed PubMed Citation Articles by Vogel, J. M. Articles by Palazzo, R. E. Pubmed/NCBI databases Substance via MeSH Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/1997//193 $5.00
The Journal of Cell Biology, Volume 137, Number 1, , 1997 193-202

Centrosomes Isolated from Spisula solidissima Oocytes Contain Rings and an Unusual Stoichiometric Ratio of /β Tubulin

Jacalyn M. Vogel^*, Tim Stearns, Conly L. Rieder, and Robert E. Palazzo^*

^* The Department of Physiology and Cell Biology, University of Kansas, Lawrence, Kansas 66045, and the Marine Biological Laboratory, Woods Hole, Massachusetts 02543; Department of Biological Sciences, Stanford University, Palo Alto, California 94305; and Wadsworth Center for Laboratories and Research, Albany, New York 12201-0509

Centrosome-dependent microtubule nucleation involves the interaction of tubulin subunits with pericentriolar material. To study the biochemical and structural basis of centrosome-dependent microtubule nucleation, centrosomes capable of organizing microtubules into astral arrays were isolated from parthenogenetically activated Spisula solidissima oocytes. Intermediate voltage electron microscopy tomography revealed that each centrosome was composed of a single centriole surrounded by pericentriolar material that was studded with ring-shaped structures 25 nm in diameter and <25 nm in length. A number of proteins copurified with centrosomes including: (a) proteins that contained M-phase–specific phosphoepitopes (MPM-2), (b) -, β-, and -tubulins, (c) actin, and (d) three low molecular weight proteins of <20 kD. -Tubulin was not an MPM-2 phosphoprotein and was the most abundant form of tubulin in centrosomes. Relatively little - or β-tubulin copurified with centrosomes, and the ratio of - to β-tubulin in centrosomes was not 1:1 as expected, but rather 1:4.6, suggesting that centrosomes contain β-tubulin that is not dimerized with -tubulin.

Abbreviations used in this paper: IVEM, intermediate voltage transmission electron microscopy; IOD, integrated optical density; MPM, M-phase– specific phosphoepitope; Mts, microtubules; PCM, pericentriolar material.

Jacalyn Vogel's current address is Department of Biology, Yale University, New Haven, CT 06511.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• DiMaio, M. A., Mikhailov, A., Rieder, C. L., Von Hoff, D. D., Palazzo, R. E. (2009). The small organic compound HMN-176 delays satisfaction of the spindle assembly checkpoint by inhibiting centrosome-dependent microtubule nucleation. Molecular Cancer Therapeutics 8: 592-601 [Abstract] [Full Text]  
• Alliegro, M. C., Alliegro, M. A., Palazzo, R. E. (2006). Centrosome-associated RNA in surf clam oocytes. Proc. Natl. Acad. Sci. USA 103: 9034-9038 [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]  
• Hannak, E., Oegema, K., Kirkham, M., Gonczy, P., Habermann, B., Hyman, A. A. (2002). The kinetically dominant assembly pathway for centrosomal asters in Caenorhabditis elegans is {gamma}-tubulin dependent. JCB 157: 591-602 [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]  
• 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]  
• Li, Q, Hansen, D, Killilea, A, Joshi, H., Palazzo, R., Balczon, R (2001). Kendrin/pericentrin-B, a centrosome protein with homology to pericentrin that complexes with PCM-1. J. Cell Sci. 114: 797-809 [Abstract]  
• 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]  
• Vogel, J, Snyder, M (2000). The carboxy terminus of Tub4p is required for gamma-tubulin function in budding yeast. J. Cell Sci. 113: 3871-3882 [Abstract]  
• Mogensen, M., Malik, A, Piel, M, Bouckson-Castaing, V, Bornens, M (2000). Microtubule minus-end anchorage at centrosomal and non-centrosomal sites: the role of ninein. J. Cell Sci. 113: 3013-3023 [Abstract]  
• 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]  
• 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. JCB 144: 721-733 [Abstract] [Full Text]  
• Wu, X., Palazzo, R. E. (1999). Differential regulation of maternal vs. paternal centrosomes. Proc. Natl. Acad. Sci. USA 96: 1397-1402 [Abstract] [Full Text]  
• Tsakraklides, V, Krogh, K, Wang, L, Bizario, J., Larson, R., Espreafico, E., Wolenski, J. (1999). Subcellular localization of GFP-myosin-V in live mouse melanocytes. J. Cell Sci. 112: 2853-2865 [Abstract]  
• Palazzo, R., Vaisberg, E., Weiss, D., Kuznetsov, S., Steffen, W (1999). Dynein is required for spindle assembly in cytoplasmic extracts of Spisula solidissima oocytes. J. Cell Sci. 112: 1291-1302 [Abstract]  
• Moritz, M., Zheng, Y., Alberts, B. M., Oegema, K. (1998). Recruitment of the {gamma}-Tubulin Ring Complex to Drosophila Salt-stripped Centrosome Scaffolds. JCB 142: 775-786 [Abstract] [Full Text]  
• Schnackenberg, B. J., Khodjakov, A., Rieder, C. L., Palazzo, R. E. (1998). The disassembly and reassembly of functional centrosomes in vitro. Proc. Natl. Acad. Sci. USA 95: 9295-9300 [Abstract] [Full Text]  
• Martin, O. C., Gunawardane, R. N., Iwamatsu, A., Zheng, Y. (1998). Xgrip109: A {gamma} Tubulin-Associated Protein with an Essential Role in {gamma} Tubulin Ring Complex ({gamma}TuRC) Assembly and Centrosome Function. JCB 141: 675-687 [Abstract] [Full Text]  
• Murphy, S. M., Urbani, L., Stearns, T. (1998). The Mammalian {gamma}-Tubulin Complex Contains Homologues of the Yeast Spindle Pole Body Components Spc97p and Spc98p. JCB 141: 663-674 [Abstract] [Full Text]  
• Dictenberg, J. B., Zimmerman, W., Sparks, C. A., Young, A., Vidair, C., Zheng, Y., Carrington, W., Fay, F. S., Doxsey, S. J. (1998). Pericentrin and {gamma}-Tubulin Form a Protein Complex and Are Organized into a Novel Lattice at the Centrosome. JCB 141: 163-174 [Abstract] [Full Text]  
• Ohta, T., Essner, R., Ryu, J.-H., Palazzo, R. E., Uetake, Y., Kuriyama, R. (2002). Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells. JCB 156: 87-100 [Abstract] [Full Text]  

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