Self-organization of a radial microtubule array by dynein-dependent nucleation of microtubules

Polarized radial arrays of cytoplasmic microtubules (MTs) with minus ends clustered at the cell center define the organization of the cytoplasm through interaction with microtubule motors bound to membrane organelles or chromosomes. It is generally assumed that the radial organization results from nucleation of MTs at the centrosome. However, radial MT array can also be attained through self-organization that requires the activity of a minus-end-directed MT motor, cytoplasmic dynein. In this study we examine the role of cytoplasmic dynein in the self-organization of a radial MT array in cytoplasmic fragments of fish melanophores lacking the centrosome. After activation of dynein motors bound to membrane-bound organelles, pigment granules, the fragments rapidly form polarized radial arrays of MTs and position pigment aggregates at their centers. We show that rearrangement of MTs in the cytoplasm is achieved through dynein-dependent MT nucleation. The radial pattern is generated by continuous disassembly and reassembly of MTs and concurrent minus-end-directed transport of pigment granules bearing the nucleation sites.

[1]  M. Kirschner,et al.  Dynamic instability of microtubule growth , 1984, Nature.

[2]  E. Salmon,et al.  How microtubules get fluorescent speckles. , 1998, Biophysical Journal.

[3]  C. Thaler,et al.  Regulation of organelle transport: Lessons from color change in fish , 1994 .

[4]  Eric Karsenti,et al.  Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts , 1996, Nature.

[5]  B. Alberts,et al.  The centrosome and cellular organization. , 1994, Annual review of biochemistry.

[6]  Vladimir Gelfand,et al.  Microtubule dynamics in fish melanophores , 1994, The Journal of cell biology.

[7]  M. McNiven,et al.  Microtubule polarity and the direction of pigment transport reverse simultaneously in surgically severed melanophore arms , 1984, Cell.

[8]  T. J. Keating,et al.  Microtubule release from the centrosome. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[9]  T. Schroer,et al.  Dynactin Is Required for Microtubule Anchoring at Centrosomes , 1999, The Journal of cell biology.

[10]  K. Ramyar,et al.  A Complex of NuMA and Cytoplasmic Dynein Is Essential for Mitotic Spindle Assembly , 1996, Cell.

[11]  Vladimir Gelfand,et al.  Kinesin is responsible for centrifugal movement of pigment granules in melanophores. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[12]  A. Hyman,et al.  Morphogenetic Properties of Microtubules and Mitotic Spindle Assembly , 1996, Cell.

[13]  M. Carlier,et al.  Hydrolysis of GTP associated with the formation of tubulin oligomers is involved in microtubule nucleation. , 1997, Biophysical journal.

[14]  T. Svitkina,et al.  Speckle microscopic evaluation of microtubule transport in growing nerve processes , 1999, Nature Cell Biology.

[15]  E. Salmon,et al.  Pathways of spindle assembly. , 1997, Current opinion in cell biology.

[16]  C. Echeverri,et al.  Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis , 1996, The Journal of cell biology.

[17]  R. Vallee,et al.  Light Intermediate Chain 1 Defines a Functional Subfraction of Cytoplasmic Dynein Which Binds to Pericentrin* , 2000, The Journal of Biological Chemistry.

[18]  Dynein, Dynactin, and Kinesin II's Interaction with Microtubules Is Regulated during Bidirectional Organelle Transport , 2000, The Journal of cell biology.

[19]  S. Tsukita,et al.  Centriolar satellites: molecular characterization, ATP-dependent movement toward centrioles and possible involvement in ciliogenesis. , 1999 .

[20]  I. Vernos,et al.  Heterotrimeric Kinesin II Is the Microtubule Motor Protein Responsible for Pigment Dispersion in Xenopus Melanophores , 1998, The Journal of cell biology.

[21]  V. Allan,et al.  Microtubule-based membrane movement. , 1998, Biochimica et biophysica acta.

[22]  K. Pfister,et al.  Differential phosphorylation in vivo of cytoplasmic dynein associated with anterogradely moving organelles , 1994, The Journal of cell biology.

[23]  D. Compton,et al.  Mitotic Spindle Poles are Organized by Structural and Motor Proteins in Addition to Centrosomes , 1997, The Journal of cell biology.

[24]  K. Kalil,et al.  Reorganization and Movement of Microtubules in Axonal Growth Cones and Developing Interstitial Branches , 1999, The Journal of Neuroscience.

[25]  T. Hyman,et al.  Recombinant p50/dynamitin as a tool to examine the role of dynactin in intracellular processes. , 1999, Methods in cell biology.

[26]  G. Borisy,et al.  Microtubule Treadmilling in Vivo , 1997, Science.

[27]  L. Goldstein,et al.  The road less traveled: emerging principles of kinesin motor utilization. , 1999, Annual review of cell and developmental biology.

[28]  S. Leibler,et al.  Self-organization of microtubules and motors , 1997, Nature.

[29]  Vladimir Gelfand,et al.  Molecular mechanisms of pigment transport in melanophores. , 1999, Pigment cell research.

[30]  Yixian Zheng,et al.  Nucleation of microtubule assembly by a γ-tubulin-containing ring complex , 1995, Nature.

[31]  D. Murphy,et al.  Purified kinesin promotes vesicle motility and induces active sliding between microtubules in vitro. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[32]  G. Borisy,et al.  Self-centring activity of cytoplasm , 1997, Nature.

[33]  E. Salmon,et al.  Actomyosin-based Retrograde Flow of Microtubules in the Lamella of Migrating Epithelial Cells Influences Microtubule Dynamic Instability and Turnover and Is Associated with Microtubule Breakage and Treadmilling , 1997, The Journal of cell biology.

[34]  Richard B. Vallee,et al.  An extended microtubule-binding structure within the dynein motor domain , 1997, Nature.

[35]  S. Doxsey,et al.  Cytoplasmic Dynein-mediated Assembly of Pericentrin and γ Tubulin onto Centrosomes , 2000 .

[36]  G. Borisy,et al.  Centrosomal control of microtubule dynamics. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[37]  G. C. Rogers,et al.  Roles of motor proteins in building microtubule-based structures: a basic principle of cellular design. , 2000, Biochimica et biophysica acta.

[38]  M. Wallin,et al.  Evidence for several roles of dynein in pigment transport in melanophores. , 1997, Cell motility and the cytoskeleton.

[39]  E. Karsenti,et al.  Taxol-induced microtubule asters in mitotic extracts of Xenopus eggs: requirement for phosphorylated factors and cytoplasmic dynein , 1991, The Journal of cell biology.

[40]  T. Schroer,et al.  Opposing motor activities are required for the organization of the mammalian mitotic spindle pole , 1996, The Journal of cell biology.

[41]  D. Compton,et al.  Spindle assembly in animal cells. , 2000, Annual review of biochemistry.

[42]  G. Borisy,et al.  Quantitative determination of the proportion of microtubule polymer present during the mitosis-interphase transition. , 1994, Journal of cell science.

[43]  C. Waterman-Storer,et al.  The p150Glued component of the dynactin complex binds to both microtubules and the actin-related protein centractin (Arp-1). , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[44]  R. Vallee,et al.  Direct Interaction of Pericentrin with Cytoplasmic Dynein Light Intermediate Chain Contributes to Mitotic Spindle Organization , 1999, The Journal of cell biology.