![[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}
|
|
|
|
|
||
Vol. 12, Issue 4, 971-980, April 2001
Tubulin
Department of Biology and Program in Molecular and Cellular
Biology, University of Massachusetts, Amherst, Massachusetts 01003
LLCPK-1 cells were transfected with a green fluorescent protein
(GFP)-
tubulin construct and a cell line permanently expressing GFP- tubulin was established (LLCPK-1). The mitotic index and doubling time for LLCPK-1 were not significantly different from parental cells. Quantitative immunoblotting showed that
17% of the tubulin in LLCPK-1 cells was GFP-tubulin; the level of
unlabeled tubulin was reduced to 82% of that in parental cells. The
parameters of microtubule dynamic instability were compared for
interphase LLCPK-1 and parental cells injected with
rhodamine-labeled tubulin. Dynamic instability was very
similar in the two cases, demonstrating that LLCPK-1 cells are a
useful tool for analysis of microtubule dynamics throughout the cell
cycle. Comparison of astral microtubule behavior in mitosis with
microtubule behavior in interphase demonstrated that the frequency of
catastrophe increased twofold and that the frequency of rescue
decreased nearly fourfold in mitotic compared with interphase cells.
The percentage of time that microtubules spent in an attenuated state,
or pause, was also dramatically reduced, from 73.5% in interphase to
11.4% in mitosis. The rates of microtubule elongation and rapid
shortening were not changed; overall dynamicity increased 3.6-fold in
mitosis. Microtubule release from the centrosome and a subset of
differentially stable astral microtubules were also observed. The
results provide the first quantitative measurements of mitotic
microtubule dynamics in mammalian cells.
Online version of this article contains video
material and is available at www.molbiocell.org.
*
Corresponding author. E-mail address: patw{at}bio.umass.edu.
This article has been cited by other articles:
|
|
 |
|
  P. J. Keller, F. Pampaloni, G. Lattanzi, and E. H. K. Stelzer Three-Dimensional Microtubule Behavior in Xenopus Egg Extracts Reveals Four Dynamic States and State-Dependent Elastic Properties Biophys. J., August 1, 2008; 95(3): 1474 - 1486. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Murthy and P. Wadsworth Dual role for microtubules in regulating cortical contractility during cytokinesis J. Cell Sci., July 15, 2008; 121(14): 2350 - 2359. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Yao, Y. Wakamatsu, T. J. Itoh, T. Shoji, and T. Hashimoto Arabidopsis SPIRAL2 promotes uninterrupted microtubule growth by suppressing the pause state of microtubule dynamics J. Cell Sci., July 15, 2008; 121(14): 2372 - 2381. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-F. Lin, S. P. Gao, D. L. Price, S. Li, T.-C. Chou, P. Singh, Y.-Y. Huang, Y. Fong, and R. J. Wong Synergy of a Herpes Oncolytic Virus and Paclitaxel for Anaplastic Thyroid Cancer Clin. Cancer Res., March 1, 2008; 14(5): 1519 - 1528. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Efimov, N. Schiefermeier, I. Grigoriev, M. C. Brown, C. E. Turner, J. V. Small, and I. Kaverina Paxillin-dependent stimulation of microtubule catastrophes at focal adhesion sites J. Cell Sci., January 15, 2008; 121(2): 196 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. P. Ferenz and P. Wadsworth Prophase Microtubule Arrays Undergo Flux-like Behavior in Mammalian Cells Mol. Biol. Cell, October 1, 2007; 18(10): 3993 - 4002. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Alli, J.-M. Yang, J. M. Ford, and W. N. Hait Reversal of Stathmin-Mediated Resistance to Paclitaxel and Vinblastine in Human Breast Carcinoma Cells Mol. Pharmacol., May 1, 2007; 71(5): 1233 - 1240. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. V. Gregoretti, G. Margolin, M. S. Alber, and H. V. Goodson Insights into cytoskeletal behavior from computational modeling of dynamic microtubules in a cell-like environment J. Cell Sci., November 15, 2006; 119(22): 4781 - 4788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Warren, A. Rutkowski, and L. Cassimeris Infection with Replication-deficient Adenovirus Induces Changes in the Dynamic Instability of Host Cell Microtubules Mol. Biol. Cell, August 1, 2006; 17(8): 3557 - 3568. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-h. Xi, F. Bai, R. McGaha, and U. P. Andley Alpha-crystallin expression affects microtubule assembly and prevents their aggregation FASEB J, May 1, 2006; 20(7): 846 - 857. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Pourroy, S. Honore, E. Pasquier, V. Bourgarel-Rey, A. Kruczynski, C. Briand, and D. Braguer Antiangiogenic concentrations of vinflunine increase the interphase microtubule dynamics and decrease the motility of endothelial cells. Cancer Res., March 15, 2006; 66(6): 3256 - 3263. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Rosa, P. Canovas, A. Islam, D. C. Altieri, and S. J. Doxsey Survivin Modulates Microtubule Dynamics and Nucleation throughout the Cell Cycle Mol. Biol. Cell, March 1, 2006; 17(3): 1483 - 1493. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Janulevicius, J. van Pelt, and A. van Ooyen Compartment Volume Influences Microtubule Dynamic Instability: A Model Study Biophys. J., February 1, 2006; 90(3): 788 - 798. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. G. O'Reilly, S. Wagner, D. J. Franks, K. Cailliau, E. Browaeys, C. Dissous, and L. A. Sabourin The Ste20-like Kinase SLK Is Required for Cell Cycle Progression through G2 J. Biol. Chem., December 23, 2005; 280(51): 42383 - 42390. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. H. Hinchcliffe Using Long-term Time-lapse Imaging of Mammalian Cell Cycle Progression for Laboratory Instruction and Analysis CBE Life Sci Educ, December 1, 2005; 4(4): 284 - 290. [Full Text] [PDF]
|
|
|
|
 |
|
 K. R. Finley and J. Berman Microtubules in Candida albicans Hyphae Drive Nuclear Dynamics and Connect Cell Cycle Progression to Morphogenesis Eukaryot. Cell, October 1, 2005; 4(10): 1697 - 1711. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Green, R. Wollman, and K. B. Kaplan APC and EB1 Function Together in Mitosis to Regulate Spindle Dynamics and Chromosome Alignment Mol. Biol. Cell, October 1, 2005; 16(10): 4609 - 4622. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. E. Kremer, T. Haystead, and I. G. Macara Mammalian Septins Regulate Microtubule Stability through Interaction with the Microtubule-binding Protein MAP4 Mol. Biol. Cell, October 1, 2005; 16(10): 4648 - 4659. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. J. Salaycik, C. J. Fagerstrom, K. Murthy, U. S. Tulu, and P. Wadsworth Quantification of microtubule nucleation, growth and dynamics in wound-edge cells J. Cell Sci., September 15, 2005; 118(18): 4113 - 4122. [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]
|
|
|
|
|
|
V. K.W. Wong, P. Chiu, S. S.M. Chung, L. M.C. Chow, Y.-Z. Zhao, B. B. Yang, and B. C.B. Ko Pseudolaric Acid B, a Novel Microtubule-Destabilizing Agent That Circumvents Multidrug Resistance Phenotype and Exhibits Antitumor Activity In vivo Clin. Cancer Res., August 15, 2005; 11(16): 6002 - 6011. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. A. Brito, J. Strauss, V. Magidson, I. Tikhonenko, A. Khodjakov, and M. P. Koonce Pushing Forces Drive the Comet-like Motility of Microtubule Arrays in Dictyostelium Mol. Biol. Cell, July 1, 2005; 16(7): 3334 - 3340. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. M. Rusan and P. Wadsworth Centrosome fragments and microtubules are transported asymmetrically away from division plane in anaphase J. Cell Biol., January 3, 2005; 168(1): 21 - 28. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhou, A. O'Brate, A. Zelnak, and P. Giannakakou Survivin Deregulation in {beta}-Tubulin Mutant Ovarian Cancer Cells Underlies Their Compromised Mitotic Response to Taxol Cancer Res., December 1, 2004; 64(23): 8708 - 8714. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Fanara, S. Turner, R. Busch, S. Killion, M. Awada, H. Turner, A. Mahsut, K. L. LaPrade, J. M. Stark, and M. K. Hellerstein In Vivo Measurement of Microtubule Dynamics Using Stable Isotope Labeling with Heavy Water: EFFECT OF TAXANES J. Biol. Chem., November 26, 2004; 279(48): 49940 - 49947. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Ohi, T. Sapra, J. Howard, and T. J. Mitchison Differentiation of Cytoplasmic and Meiotic Spindle Assembly MCAK Functions by Aurora B-dependent Phosphorylation Mol. Biol. Cell, June 1, 2004; 15(6): 2895 - 2906. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Paluh, A. N. Killilea, H. W. Detrich 3rd, and K. H. Downing Meiosis-specific Failure of Cell Cycle Progression in Fission Yeast by Mutation of a Conserved {beta}-Tubulin Residue Mol. Biol. Cell, March 1, 2004; 15(3): 1160 - 1171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Sagolla, S. Uzawa, and W. Z. Cande Individual microtubule dynamics contribute to the function of mitotic and cytoplasmic arrays in fission yeast J. Cell Sci., December 15, 2003; 116(24): 4891 - 4903. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kamath and M. A. Jordan Suppression of Microtubule Dynamics by Epothilone B Is Associated with Mitotic Arrest Cancer Res., September 15, 2003; 63(18): 6026 - 6031. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. L. Tinley, D. A. Randall-Hlubek, R. M. Leal, E. M. Jackson, J. W. Cessac, J. C. Quada Jr., T. K. Hemscheidt, and S. L. Mooberry Taccalonolides E and A: Plant-derived Steroids with Microtubule-stabilizing Activity Cancer Res., June 15, 2003; 63(12): 3211 - 3220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kelling, K. Sullivan, L. Wilson, and M. A. Jordan Suppression of Centromere Dynamics by Taxol(R) in Living Osteosarcoma Cells Cancer Res., June 1, 2003; 63(11): 2794 - 2801. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. M.J. Hut, W. Lemstra, E. H. Blaauw, G. W.A. van Cappellen, H. H. Kampinga, and O. C.M. Sibon Centrosomes Split in the Presence of Impaired DNA Integrity during Mitosis Mol. Biol. Cell, May 1, 2003; 14(5): 1993 - 2004. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Shirasu-Hiza, P. Coughlin, and T. Mitchison Identification of XMAP215 as a microtubule-destabilizing factor in Xenopus egg extract by biochemical purification J. Cell Biol., April 28, 2003; 161(2): 349 - 358. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. L. Tinley, R. M. Leal, D. A. Randall-Hlubek, J. W. Cessac, L. R. Wilkens, P. N. Rao, and S. L. Mooberry Novel 2-Methoxyestradiol Analogues with Antitumor Activity Cancer Res., April 1, 2003; 63(7): 1538 - 1549. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Schmoranzer and S. M. Simon Role of Microtubules in Fusion of Post-Golgi Vesicles to the Plasma Membrane Mol. Biol. Cell, April 1, 2003; 14(4): 1558 - 1569. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Dhonukshe and T. W. J. Gadella Jr. Alteration of Microtubule Dynamic Instability during Preprophase Band Formation Revealed by Yellow Fluorescent Protein-CLIP170 Microtubule Plus-End Labeling PLANT CELL, March 1, 2003; 15(3): 597 - 611. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Piehl and L. Cassimeris Organization and Dynamics of Growing Microtubule Plus Ends during Early Mitosis Mol. Biol. Cell, March 1, 2003; 14(3): 916 - 925. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Gergely, V. M. Draviam, and J. W. Raff The ch-TOG/XMAP215 protein is essential for spindle pole organization in human somatic cells Genes & Dev., February 1, 2003; 17(3): 336 - 341. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Tirnauer, J. C. Canman, E.D. Salmon, and T. J. Mitchison EB1 Targets to Kinetochores with Attached, Polymerizing Microtubules Mol. Biol. Cell, December 1, 2002; 13(12): 4308 - 4316. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Tirnauer, S. Grego, E.D. Salmon, and T. J. Mitchison EB1-Microtubule Interactions in Xenopus Egg Extracts: Role of EB1 in Microtubule Stabilization and Mechanisms of Targeting to Microtubules Mol. Biol. Cell, October 1, 2002; 13(10): 3614 - 3626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. M. Rusan, U. S. Tulu, C. Fagerstrom, and P. Wadsworth Reorganization of the microtubule array in prophase/prometaphase requires cytoplasmic dynein-dependent microtubule transport J. Cell Biol., September 16, 2002; 158(6): 997 - 1003. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Rogers, G. C. Rogers, D. J. Sharp, and R. D. Vale Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle J. Cell Biol., September 3, 2002; 158(5): 873 - 884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Kline-Smith and C. E. Walczak The Microtubule-destabilizing Kinesin XKCM1 Regulates Microtubule Dynamic Instability in Cells Mol. Biol. Cell, August 1, 2002; 13(8): 2718 - 2731. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. W. Landen, R. Lang, S. J. McMahon, N. M. Rusan, A.-M. Yvon, A. W. Adams, M. D. Sorcinelli, R. Campbell, P. Bonaccorsi, J. C. Ansel, et al. Noscapine Alters Microtubule Dynamics in Living Cells and Inhibits the Progression of Melanoma Cancer Res., July 15, 2002; 62(14): 4109 - 4114. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Joglekar and A. J. Hunt A Simple, Mechanistic Model for Directional Instability during Mitotic Chromosome Movements Biophys. J., July 1, 2002; 83(1): 42 - 58. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Buster, K. McNally, and F. J. McNally Katanin inhibition prevents the redistribution of {gamma}-tubulin at mitosis J. Cell Sci., January 3, 2002; 115(5): 1083 - 1092. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Kinoshita, I. Arnal, A. Desai, D. N. Drechsel, and A. A. Hyman Reconstitution of Physiological Microtubule Dynamics Using Purified Components Science, November 9, 2001; 294(5545): 1340 - 1343. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Goncalves, D. Braguer, K. Kamath, L. Martello, C. Briand, S. Horwitz, L. Wilson, and M. A. Jordan Resistance to Taxol in lung cancer cells associated with increased microtubule dynamics PNAS, September 13, 2001; (2001) 191388598. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-M. C. Yvon, D. J. Gross, and P. Wadsworth Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells PNAS, June 28, 2001; (2001) 141224198. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Eichenmuller, P. Everley, J. Palange, D. Lepley, and K. A. Suprenant The Human EMAP-like Protein-70 (ELP70) Is a Microtubule Destabilizer That Localizes to the Mitotic Apparatus J. Biol. Chem., January 4, 2002; 277(2): 1301 - 1309. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-M. C. Yvon, D. J. Gross, and P. Wadsworth Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells PNAS, July 17, 2001; 98(15): 8656 - 8661. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Goncalves, D. Braguer, K. Kamath, L. Martello, C. Briand, S. Horwitz, L. Wilson, and M. A. Jordan Resistance to Taxol in lung cancer cells associated with increased microtubule dynamics PNAS, September 25, 2001; 98(20): 11737 - 11742. [Abstract] [Full Text] [PDF]
|
|
