Journal of Cell Science, Vol 95, Issue 3 405-411, Copyright © 1990 by Company of Biologists

JOURNAL ARTICLES

Nucleation of microtubules from mitotic centrosomes is modulated by a phosphorylated epitope

VE Centonze and GG Borisy
Laboratory of Molecular Biology, University of Wisconsin-Madison 53706.

At the onset of mitosis a class of proteins appears that possess a phosphorylated epitope recognized by the monoclonal antibody MPM-2. Immunofluorescence staining shows that a subset of these proteins is associated with the centrosomes of the mitotic apparatus. The appearance of these proteins coincides with the increased microtubule nucleating capacity of the centrosomes. We have tested whether growth of microtubules from mitotic centrosomes in a lysed cell model is dependent on the availability of the phosphorylated epitope by blocking the epitope with a specific antibody or by modifying it by removal of the phosphate. Centrosomes incubated with purified tubulin nucleate microtubule asters. However, preincubating the centrosomes with MPM-2 blocks all microtubule nucleation. Pretreating mitotic centrosomes with alkaline phosphatase also inhibits nucleation. These data suggest that the phosphorylated epitope recognized by MPM-2 is important for microtubule nucleation.

This article has been cited by other articles:

				S. Sankaran, D. E. Crone, R. E. Palazzo, and J. D. Parvin Aurora-A Kinase Regulates Breast Cancer Associated Gene 1 Inhibition of Centrosome-Dependent Microtubule Nucleation Cancer Res., December 1, 2007; 67(23): 11186 - 11194. [Abstract] [Full Text] [PDF]

				A. Straube, M. Brill, B. R. Oakley, T. Horio, and G. Steinberg Microtubule Organization Requires Cell Cycle-dependent Nucleation at Dispersed Cytoplasmic Sites: Polar and Perinuclear Microtubule Organizing Centers in the Plant Pathogen Ustilago maydis Mol. Biol. Cell, February 1, 2003; 14(2): 642 - 657. [Abstract] [Full Text] [PDF]

				L. S. Kiat, K. M. Hui, and G. Gopalan Aurora-A Kinase Interacting Protein (AIP), a Novel Negative Regulator of Human Aurora-A Kinase J. Biol. Chem., November 15, 2002; 277(47): 45558 - 45565. [Abstract] [Full Text] [PDF]

				Q. Lu, R. L. Dunn, R. Angeles, and G. D. Smith Regulation of Spindle Formation by Active Mitogen-Activated Protein Kinase and Protein Phosphatase 2A During Mouse Oocyte Meiosis Biol Reprod, January 1, 2002; 66(1): 29 - 37. [Abstract] [Full Text]

				A. Fry, P Descombes, C Twomey, R Bacchieri, and E. Nigg The NIMA-related kinase X-Nek2B is required for efficient assembly of the zygotic centrosome in Xenopus laevis J. Cell Sci., January 6, 2000; 113(11): 1973 - 1984. [Abstract] [PDF]

				A. Khodjakov and C. L. Rieder The Sudden Recruitment of {gamma}-Tubulin to the Centrosome at the Onset of Mitosis and its Dynamic Exchange Throughout the Cell Cycle, Do Not Require Microtubules J. Cell Biol., August 9, 1999; 146(3): 585 - 596. [Abstract] [Full Text] [PDF]

				R Giet and C Prigent Aurora/Ipl1p-related kinases, a new oncogenic family of mitotic serine-threonine kinases J. Cell Sci., January 11, 1999; 112(21): 3591 - 3601. [Abstract] [PDF]

				A. M. Fry, T. Mayor, P. Meraldi, Y.-D. Stierhof, K. Tanaka, and E. A. Nigg C-Nap1, a Novel Centrosomal Coiled-Coil Protein and Candidate Substrate of the Cell Cycle-regulated Protein Kinase Nek2 J. Cell Biol., June 29, 1998; 141(7): 1563 - 1574. [Abstract] [Full Text] [PDF]

				E Logarinho and C. Sunkel The Drosophila POLO kinase localises to multiple compartments of the mitotic apparatus and is required for the phosphorylation of MPM2 reactive epitopes J. Cell Sci., January 10, 1998; 111(19): 2897 - 2909. [Abstract] [PDF]

				N. Helps, N. Brewis, K Lineruth, T Davis, K Kaiser, and P. Cohen Protein phosphatase 4 is an essential enzyme required for organisation of microtubules at centrosomes in Drosophila embryos J. Cell Sci., January 5, 1998; 111(10): 1331 - 1340. [Abstract] [PDF]

				C Roghi, R Giet, R Uzbekov, N Morin, I Chartrain, R Le Guellec, A Couturier, M Doree, M Philippe, and C Prigent The Xenopus protein kinase pEg2 associates with the centrosome in a cell cycle-dependent manner, binds to the spindle microtubules and is involved in bipolar mitotic spindle assembly J. Cell Sci., January 3, 1998; 111(5): 557 - 572. [Abstract] [PDF]

				M. Kimura, S. Kotani, T. Hattori, N. Sumi, T. Yoshioka, K. Todokoro, and Y. Okano Cell Cycle-dependent Expression and Spindle Pole Localization of a Novel Human Protein Kinase, Aik, Related to Aurora of Drosophila and Yeast Ipl1 J. Biol. Chem., May 23, 1997; 272(21): 13766 - 13771. [Abstract] [Full Text] [PDF]

				A. Tassin, C Celati, M Paintrand, and M Bornens Identification of an Spc110p-related protein in vertebrates J. Cell Sci., January 10, 1997; 110(20): 2533 - 2545. [Abstract] [PDF]

				K Oegema, W. Marshall, J. Sedat, and B. Alberts Two proteins that cycle asynchronously between centrosomes and nuclear structures: Drosophila CP60 and CP190 J. Cell Sci., January 7, 1997; 110(14): 1573 - 1583. [Abstract] [PDF]

				M. Martin, S. Osmani, and B. Oakley The role of gamma-tubulin in mitotic spindle formation and cell cycle progression in Aspergillus nidulans J. Cell Sci., January 3, 1997; 110(5): 623 - 633. [Abstract] [PDF]

				H Masuda and T Shibata Role of gamma-tubulin in mitosis-specific microtubule nucleation from the Schizosaccharomyces pombe spindle pole body J. Cell Sci., January 1, 1996; 109(1): 165 - 177. [Abstract] [PDF]

				F. Ahmad and P. Baas Microtubules released from the neuronal centrosome are transported into the axon J. Cell Sci., January 8, 1995; 108(8): 2761 - 2769. [Abstract] [PDF]

				R. Burns Analysis of the gamma-tubulin sequences: implications for the functional properties of gamma-tubulin J. Cell Sci., January 6, 1995; 108(6): 2123 - 2130. [PDF]

				M E Lane and D Kalderon RNA localization along the anteroposterior axis of the Drosophila oocyte requires PKA-mediated signal transduction to direct normal microtubule organization. Genes & Dev., December 15, 1994; 8(24): 2986 - 2995. [Abstract] [PDF]

				J. Dominguez, B Buendia, C Lopez-Otin, C Antony, E Karsenti, and J Avila A protein related to brain microtubule-associated protein MAP1B is a component of the mammalian centrosome J. Cell Sci., January 2, 1994; 107(2): 601 - 611. [Abstract] [PDF]

				E. Paul and A Quaroni Identification of a 102 kDa protein (cytocentrin) immunologically related to keratin 19, which is a cytoplasmically derived component of the mitotic spindle pole J. Cell Sci., January 11, 1993; 106(3): 967 - 981. [Abstract] [PDF]

				K Ohta, N Shiina, E Okumura, S Hisanaga, T Kishimoto, S Endo, Y Gotoh, E Nishida, and H Sakai Microtubule nucleating activity of centrosomes in cell-free extracts from Xenopus eggs: involvement of phosphorylation and accumulation of pericentriolar material J. Cell Sci., January 1, 1993; 104(1): 125 - 137. [Abstract] [PDF]

				C.E. Alfa, I.M. Gallagher, and J.S. Hyams Subcellular Localization of the p34cdc2/p63cdc13 Protein Kinase in Fission Yeast Cold Spring Harb Symp Quant Biol, January 1, 1991; 56(0): 489 - 494. [Abstract] [PDF]

				B. Buendia, P.R. Clarke, M.A. Felix, E. Karsenti, D. Leiss, and F. Verde Regulation of Protein Kinases Associated with Cyclin A and Cyclin B and Their Effect on Microtubule Dynamics and Nucleation in Xenopus Egg Extracts Cold Spring Harb Symp Quant Biol, January 1, 1991; 56(0): 523 - 532. [Abstract] [PDF]

				D. B. Friedman, J. W. Kern, B. J. Huneycutt, D. B. N. Vinh, D. K. Crawford, E. Steiner, D. Scheiltz, J. Yates III, K. A. Resing, N. G. Ahn, et al. Yeast Mps1p Phosphorylates the Spindle Pole Component Spc110p in the N-terminal Domain J. Biol. Chem., May 18, 2001; 276(21): 17958 - 17967. [Abstract] [Full Text] [PDF]

				W. Lutz, W. L. Lingle, D. McCormick, T. M. Greenwood, and J. L. Salisbury Phosphorylation of Centrin during the Cell Cycle and Its Role in Centriole Separation Preceding Centrosome Duplication J. Biol. Chem., June 1, 2001; 276(23): 20774 - 20780. [Abstract] [Full Text] [PDF]