|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Research Article |
1 Department of Cell and Molecular Biology, Northwestern University Medical
School, Chicago, IL 60611, USA
2 Laboratory of Cell Motility, A. N. Belozersky Institute, Moscow State
University, Moscow, Russia
* Author for correspondence (e-mail: y-komarova{at}northwestern.edu)
Accepted 20 June 2002
Microtubule dynamics were investigated in CHO and NRK cells by novel experimental approaches designed to evaluate the microtubule behavior in the cell interior. These approaches were: (1) laser photobleaching of a path through the centrosome; (2) direct observation of microtubules in centrosome-containing cytoplasts; (3) GFP-CLIP-170 expression as a marker for microtubule plus end growth; and (iv) sequential subtraction analysis. The combination of these approaches allowed us to obtain data where the density of microtubules had previously prevented conventional methods to be applicable.
In the steady state, nascent microtubules grew persistently from the centrosome towards the cell margin. Frequently, they arrived at the cell margin without undergoing any transition to the shortening phase. In contrast to the growth of microtubules, shortening of the plus ends from the periphery was non-persistent; that is, rescue was frequent and the extent of shortening showed a distribution of lengths reflecting a stochastic process. The combination of persistent growth and a cell boundary led to a difference in apparent microtubule behavior in the cell interior compared with that near the cell margin. Whereas microtubules in the cell interior showed asymmetric transition frequencies, their behavior near the cell margin showed frequent fluctuations between phases of shortening and growth. Complete microtubule turnover was accomplished by the relatively rare episodes of shortening back to the centrosome. Release from the centrosome with subsequent minus end shortening also occurred but was a minor mechanism for microtubule turnover compared with the plus end pathway.
We propose a life cycle for a microtubule which consists of rapid growth from the centrosome to the cell margin followed by an indefinite period of fluctuations of phases of shortening and growth. We suggest that persistent growth and asymmetric transition frequencies serve the biological function of providing a mechanism by which microtubules may rapidly accommodate to the changing shape and advancing edge of motile cells.
Key words: Microtubules, Dynamics, Mammalian cell culture line
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
Related articles in JCS:
This article has been cited by other articles:
|
|
 |
|
  V. Daire, J. Giustiniani, I. Leroy-Gori, M. Quesnoit, S. Drevensek, A. Dimitrov, F. Perez, and C. Pous Kinesin-1 Regulates Microtubule Dynamics via a c-Jun N-terminal Kinase-dependent Mechanism J. Biol. Chem., November 13, 2009; 284(46): 31992 - 32001. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Komarova, C. O. De Groot, I. Grigoriev, S. M. Gouveia, E. L. Munteanu, J. M. Schober, S. Honnappa, R. M. Buey, C. C. Hoogenraad, M. Dogterom, et al. Mammalian end binding proteins control persistent microtubule growth J. Cell Biol., March 9, 2009; 184(5): 691 - 706. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. A. Komarova, D. Mehta, and A. B. Malik Dual Regulation of Endothelial Junctional Permeability Sci. Signal., November 13, 2007; 2007(412): re8 - re8. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. P. Brangwynne, F. C. MacKintosh, and D. A. Weitz Force fluctuations and polymerization dynamics of intracellular microtubules PNAS, October 9, 2007; 104(41): 16128 - 16133. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Schober, Y. A. Komarova, O. Y. Chaga, A. Akhmanova, and G. G. Borisy Microtubule-targeting-dependent reorganization of filopodia J. Cell Sci., April 1, 2007; 120(7): 1235 - 1244. [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]
|
|
|
|
 |
|
 V. P. Efimov, J. Zhang, and X. Xiang CLIP-170 Homologue and NUDE Play Overlapping Roles in NUDF Localization in Aspergillus nidulans Mol. Biol. Cell, April 1, 2006; 17(4): 2021 - 2034. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Xiang A +TIP for a smooth trip J. Cell Biol., February 27, 2006; 172(5): 651 - 654. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Reilein, S. Yamada, and W. J. Nelson Self-organization of an acentrosomal microtubule network at the basal cortex of polarized epithelial cells J. Cell Biol., December 5, 2005; 171(5): 845 - 855. [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]
|
|
|
|
|
|
T. Wittmann and C. M. Waterman-Storer Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3{beta} in migrating epithelial cells J. Cell Biol., June 20, 2005; 169(6): 929 - 939. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. BALUSKA, D. VOLKMANN, and P. W. BARLOW Eukaryotic Cells and their Cell Bodies: Cell Theory Revised Ann. Bot., July 1, 2004; 94(1): 9 - 32. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Piehl, U. S. Tulu, P. Wadsworth, and L. Cassimeris Centrosome maturation: Measurement of microtubule nucleation throughout the cell cycle by using GFP-tagged EB1 PNAS, February 10, 2004; 101(6): 1584 - 1588. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D'Addario, P. D. Arora, R. P. Ellen, and C. A. G. McCulloch Regulation of Tension-induced Mechanotranscriptional Signals by the Microtubule Network in Fibroblasts J. Biol. Chem., December 26, 2003; 278(52): 53090 - 53097. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Green and K. B. Kaplan Chromosome instability in colorectal tumor cells is associated with defects in microtubule plus-end attachments caused by a dominant mutation in APC J. Cell Biol., December 8, 2003; 163(5): 949 - 961. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Janson, M. E. de Dood, and M. Dogterom Dynamic instability of microtubules is regulated by force J. Cell Biol., June 23, 2003; 161(6): 1029 - 1034. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Stepanova, J. Slemmer, C. C. Hoogenraad, G. Lansbergen, B. Dortland, C. I. De Zeeuw, F. Grosveld, G. van Cappellen, A. Akhmanova, and N. Galjart Visualization of Microtubule Growth in Cultured Neurons via the Use of EB3-GFP (End-Binding Protein 3-Green Fluorescent Protein) J. Neurosci., April 1, 2003; 23(7): 2655 - 2664. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Abal, M. Piel, V. Bouckson-Castaing, M. Mogensen, J.-B. Sibarita, and M. Bornens Microtubule release from the centrosome in migrating cells J. Cell Biol., December 9, 2002; 159(5): 731 - 737. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. A. Komarova, A. S. Akhmanova, S.-i. Kojima, N. Galjart, and G. G. Borisy Cytoplasmic linker proteins promote microtubule rescue in vivo J. Cell Biol., November 25, 2002; 159(4): 589 - 599. [Abstract] [Full Text] [PDF]
|
|
