![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
JC Waters, TJ Mitchison, CL Rieder and ED Salmon
Department of Biology, University of North Carolina, Chapel Hill 27599- 3280, USA.
During metaphase and anaphase in newt lung cells, tubulin subunits within the kinetochore microtubule (kMT) lattice flux slowly poleward as kMTs depolymerize at their minus-ends within in the pole. Very little is known about how and where the force that moves the tubulin subunits poleward is generated and what function it serves during mitosis. We found that treatment with the drug taxol (10 microM) caused separated centrosomes in metaphase newt lung cells to move toward one another with an average velocity of 0.89 microns/min, until the interpolar distance was reduced by 22-62%. This taxol-induced spindle shortening occurred as kMTs between the chromosomes and the poles shortened. Photoactivation of fluorescent marks on kMTs revealed that taxol inhibited kinetochore microtubule assembly/disassembly at kinetochores, whereas minus-end MT disassembly continued at a rate typical of poleward flux in untreated metaphase cells. This poleward flux was strong enough to stretch the centromeric chromatin between sister kinetochores as much as it is stretched in control metaphase cells. In anaphase, taxol blocked kMT disassembly/assembly at the kinetochore whereas minus-end disassembly continued at a rate similar to flux in control cells (approximately 0.2 microns/min). These results reveal that the mechanism for kMT poleward flux 1) is not dependent on kMT plus-end dynamics and 2) produces pulling forces capable of generating tension across the centromeres of bioriented chromosomes.
This article has been cited by other articles:
|
|
 |
|
  S. A. Ribeiro, J. C. Gatlin, Y. Dong, A. Joglekar, L. Cameron, D. F. Hudson, C. J. Farr, B. F. McEwen, E. D. Salmon, W. C. Earnshaw, et al. Condensin Regulates the Stiffness of Vertebrate Centromeres Mol. Biol. Cell, May 1, 2009; 20(9): 2371 - 2380. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Toso, J. R. Winter, A. J. Garrod, A. C. Amaro, P. Meraldi, and A. D. McAinsh Kinetochore-generated pushing forces separate centrosomes during bipolar spindle assembly J. Cell Biol., February 9, 2009; 184(3): 365 - 372. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. W. Buster, D. Zhang, and D. J. Sharp Poleward Tubulin Flux in Spindles: Regulation and Function in Mitotic Cells Mol. Biol. Cell, August 1, 2007; 18(8): 3094 - 3104. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T.-T B. Ngo, T. Peng, X.-J. Liang, O. Akeju, S. Pastorino, W. Zhang, Y. Kotliarov, J. C. Zenklusen, H. A. Fine, D. Maric, et al. The 1p-Encoded Protein Stathmin and Resistance of Malignant Gliomas to Nitrosoureas J Natl Cancer Inst, April 18, 2007; 99(8): 639 - 652. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. McNally, A. Audhya, K. Oegema, and F. J. McNally Katanin controls mitotic and meiotic spindle length J. Cell Biol., December 18, 2006; 175(6): 881 - 891. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Dhonukshe, N. Vischer, and T. W. J. Gadella Jr Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells. J. Cell Sci., August 1, 2006; 119(Pt 15): 3193 - 3205. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. D. Dao, A. Duffy, B. A. Van Tine, S.-Y. Wu, C.-M. Chiang, T. R. Broker, and L. T. Chow Dynamic Localization of the Human Papillomavirus Type 11 Origin Binding Protein E2 through Mitosis While in Association with the Spindle Apparatus. J. Virol., May 1, 2006; 80(10): 4792 - 4800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. A. Cameron, G. Yang, D. Cimini, J. C. Canman, O. Kisurina-Evgenieva, A. Khodjakov, G. Danuser, and E.D. Salmon Kinesin 5-independent poleward flux of kinetochore microtubules in PtK1 cells J. Cell Biol., April 24, 2006; 173(2): 173 - 179. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Tokai-Nishizumi, M. Ohsugi, E. Suzuki, and T. Yamamoto The Chromokinesin Kid Is Required for Maintenance of Proper Metaphase Spindle Size Mol. Biol. Cell, November 1, 2005; 16(11): 5455 - 5463. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. B. Shannon, J. C. Canman, C. B. Moree, J. S. Tirnauer, and E. D. Salmon Taxol-stabilized Microtubules Can Position the Cytokinetic Furrow in Mammalian Cells Mol. Biol. Cell, September 1, 2005; 16(9): 4423 - 4436. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. L. Moore, G. Stanvitch, M. B. Roth, and D. Rosen HCP-4/CENP-C Promotes the Prophase Timing of Centromere Resolution by Enabling the Centromere Association of HCP-6 in Caenorhabditis elegans Mol. Cell. Biol., April 1, 2005; 25(7): 2583 - 2592. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. C. Rogers, S. L. Rogers, and D. J. Sharp Spindle microtubules in flux J. Cell Sci., March 15, 2005; 118(6): 1105 - 1116. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. LaFountain Jr., C. S. Cohan, A. J. Siegel, and D. J. LaFountain Direct Visualization of Microtubule Flux during Metaphase and Anaphase in Crane-Fly Spermatocytes Mol. Biol. Cell, December 1, 2004; 15(12): 5724 - 5732. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-C. Labbe, E. K. McCarthy, and B. Goldstein The forces that position a mitotic spindle asymmetrically are tethered until after the time of spindle assembly J. Cell Biol., October 25, 2004; 167(2): 245 - 256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Gaetz and T. M. Kapoor Dynein/dynactin regulate metaphase spindle length by targeting depolymerizing activities to spindle poles J. Cell Biol., August 16, 2004; 166(4): 465 - 471. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. A. Beardmore, L. J. Ahonen, G. J. Gorbsky, and M. J. Kallio Survivin dynamics increases at centromeres during G2/M phase transition and is regulated by microtubule-attachment and Aurora B kinase activity J. Cell Sci., August 15, 2004; 117(18): 4033 - 4042. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.-Y. Hung, H.-L. Chen, C.-W. Chang, B.-R. Li, and T. K. Tang Identification of a Novel Microtubule-destabilizing Motif in CPAP That Binds to Tubulin Heterodimers and Inhibits Microtubule Assembly Mol. Biol. Cell, June 1, 2004; 15(6): 2697 - 2706. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Kline-Smith, A. Khodjakov, P. Hergert, and C. E. Walczak Depletion of Centromeric MCAK Leads to Chromosome Congression and Segregation Defects Due to Improper Kinetochore Attachments Mol. Biol. Cell, March 1, 2004; 15(3): 1146 - 1159. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Levesque, L. Howard, M. B. Gordon, and D. A. Compton A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells Mol. Biol. Cell, September 1, 2003; 14(9): 3541 - 3552. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Maddox, A. Straight, P. Coughlin, T. J. Mitchison, and E. D. Salmon Direct observation of microtubule dynamics at kinetochores in Xenopus extract spindles: implications for spindle mechanics J. Cell Biol., August 4, 2003; 162(3): 377 - 382. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Brust-Mascher and J. M. Scholey Microtubule Flux and Sliding in Mitotic Spindles of Drosophila Embryos Mol. Biol. Cell, November 1, 2002; 13(11): 3967 - 3975. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhou, J. Yao, and H. C. Joshi Attachment and tension in the spindle assembly checkpoint J. Cell Sci., September 15, 2002; 115(18): 3547 - 3555. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Kapoor and D. A. Compton Searching for the middle ground: mechanisms of chromosome alignment during mitosis J. Cell Biol., May 13, 2002; 157(4): 551 - 556. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. LaFountain Jr., R. Oldenbourg, R. W. Cole, and C. L. Rieder Microtubule Flux Mediates Poleward Motion of Acentric Chromosome Fragments during Meiosis in Insect Spermatocytes Mol. Biol. Cell, December 1, 2001; 12(12): 4054 - 4065. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Levesque and D. A. Compton The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles J. Cell Biol., September 17, 2001; 154(6): 1135 - 1146. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. K. Ngan, K. Bellman, B. T. Hill, L. Wilson, and M. A. Jordan Mechanism of Mitotic Block and Inhibition of Cell Proliferation by the Semisynthetic Vinca Alkaloids Vinorelbine and Its Newer Derivative Vinflunine Mol. Pharmacol., July 1, 2001; 60(1): 225 - 232. [Abstract] [Full Text]
|
|
|
|
|
|
D. B. Hoffman, C. G. Pearson, T. J. Yen, B. J. Howell, and E.D. Salmon Microtubule-dependent Changes in Assembly of Microtubule Motor Proteins and Mitotic Spindle Checkpoint Proteins at PtK1 Kinetochores Mol. Biol. Cell, July 1, 2001; 12(7): 1995 - 2009. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. G. Pearson, P. S. Maddox, E.D. Salmon, and K. Bloom Budding Yeast Chromosome Structure and Dynamics during Mitosis J. Cell Biol., March 19, 2001; 152(6): 1255 - 1266. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. B. Gordon, L. Howard, and D. A. Compton Chromosome Movement in Mitosis Requires Microtubule Anchorage at Spindle Poles J. Cell Biol., January 29, 2001; 152(3): 425 - 434. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. B. Nicklas, J. C. Waters, E. D. Salmon, and S. C. Ward Checkpoint signals in grasshopper meiosis are sensitive to microtubule attachment, but tension is still essential J. Cell Sci., January 12, 2001; 114(23): 4173 - 4183. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B.J. Howell, D.B. Hoffman, G. Fang, A.W. Murray, and E.D. Salmon Visualization of Mad2 Dynamics at Kinetochores, along Spindle Fibers, and at Spindle Poles in Living Cells J. Cell Biol., September 18, 2000; 150(6): 1233 - 1250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Waterman-Storer, W. C. Salmon, and E.D. Salmon Feedback Interactions between Cell-Cell Adherens Junctions and Cytoskeletal Dynamics in Newt Lung Epithelial Cells Mol. Biol. Cell, July 1, 2000; 11(7): 2471 - 2483. [Abstract] [Full Text]
|
|
|
|
|
|
A. Hunter and L Wordeman How motor proteins influence microtubule polymerization dynamics J. Cell Sci., January 12, 2000; 113(24): 4379 - 4389. [Abstract] [PDF]
|
|
|
|
|
|
L Quarmby Cellular Samurai: katanin and the severing of microtubules J. Cell Sci., January 8, 2000; 113(16): 2821 - 2827. [Abstract] [PDF]
|
|
|
|
|
|
K. McNally, O. Bazirgan, and F. McNally Two domains of p80 katanin regulate microtubule severing and spindle pole targeting by p60 katanin J. Cell Sci., January 5, 2000; 113(9): 1623 - 1633. [Abstract] [PDF]
|
|
|
|
|
|
K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, and M. Yanagida Dynamics of Centromeres during Metaphase-Anaphase Transition in Fission Yeast: Dis1 Is Implicated in Force Balance in Metaphase Bipolar Spindle Mol. Biol. Cell, November 1, 1998; 9(11): 3211 - 3225. [Abstract] [Full Text]
|
|
|
|
|
|
S. Bonaccorsi, M. G. Giansanti, and M. Gatti Spindle Self-organization and Cytokinesis During Male Meiosis in asterless Mutants of Drosophila melanogaster J. Cell Biol., August 10, 1998; 142(3): 751 - 761. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. J. McNally and S. Thomas Katanin Is Responsible for the M-Phase Microtubule-severing Activity in Xenopus Eggs Mol. Biol. Cell, July 1, 1998; 9(7): 1847 - 1861. [Abstract] [Full Text]
|
|
|
|
|
|
M. Kallio, J. Weinstein, J. R. Daum, D. J. Burke, and G. J. Gorbsky Mammalian p55CDC Mediates Association of the Spindle Checkpoint Protein Mad2 with the Cyclosome/Anaphase-promoting Complex, and is Involved in Regulating Anaphase Onset and Late Mitotic Events J. Cell Biol., June 15, 1998; 141(6): 1393 - 1406. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Waters, R.-H. Chen, A. W. Murray, and E.D. Salmon Localization of Mad2 to Kinetochores Depends on Microtubule Attachment, Not Tension J. Cell Biol., June 1, 1998; 141(5): 1181 - 1191. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Desai, P. S. Maddox, T. J. Mitchison, and E.D. Salmon Anaphase A Chromosome Movement and Poleward Spindle Microtubule Flux Occur At Similar Rates in Xenopus Extract Spindles J. Cell Biol., May 4, 1998; 141(3): 703 - 713. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-G. Yu, E. N. Hiatt, A. Chan, M. Sweeney, and R. K. Dawe Neocentromere-mediated Chromosome Movement in Maize J. Cell Biol., November 17, 1997; 139(4): 831 - 840. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Waterman-Storer and E.D. Salmon 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 J. Cell Biol., October 20, 1997; 139(2): 417 - 434. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Gaglio, M. A. Dionne, and D. A. Compton Mitotic Spindle Poles are Organized by Structural and Motor Proteins in Addition to Centrosomes J. Cell Biol., September 8, 1997; 138(5): 1055 - 1066. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Wheatley, E. H. Hinchcliffe, M. Glotzer, A. A. Hyman, G. Sluder, and Y.-l. Wang CDK1 Inactivation Regulates Anaphase Spindle Dynamics and Cytokinesis In Vivo J. Cell Biol., July 28, 1997; 138(2): 385 - 393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. F. McEwen, A. B. Heagle, G. O. Cassels, K. F. Buttle, and C. L. Rieder Kinetochore Fiber Maturation in PtK1 Cells and Its Implications for the Mechanisms of Chromosome Congression and Anaphase Onset J. Cell Biol., June 30, 1997; 137(7): 1567 - 1580. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Waters, R. Skibbens, and E. Salmon Oscillating mitotic newt lung cell kinetochores are, on average, under tension and rarely push J. Cell Sci., January 12, 1996; 109(12): 2823 - 2831. [Abstract] [PDF]
|
|
|
|
 |
|
 E. N. Hiatt, E. K. Kentner, and R. K. Dawe Independently Regulated Neocentromere Activity of Two Classes of Tandem Repeat Arrays PLANT CELL, February 1, 2002; 14(2): 407 - 420. [Abstract] [Full Text] [PDF]
|
|
