Microtubule dynamics in vivo: a test of mechanisms of turnover

doi:10.1083/jcb.104.3.395

This Article

	Full Text (PDF, 3714K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Sammak, P. J.
	Articles by Borisy, G. G.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Sammak, P. J.
	Articles by Borisy, G. G.


Social Bookmarking

	What's this?

ARTICLES

Microtubule dynamics in vivo: a test of mechanisms of turnover

PJ Sammak, GJ Gorbsky and GG Borisy

Clarification of the mechanism of microtubule dynamics requires an analysis of the microtubule pattern at two time points in the same cell with single fiber resolution. Single microtubule resolution was obtained by microinjection of haptenized tubulin (fluorescein-tubulin) and subsequent indirect immunofluorescence with an antifluorescein antibody. The two time points in a single cell were, first, the time of photobleaching fluorescein-tubulin, and second, the time of fixation. The pattern of fluorescence replacement in the bleached zone during this time interval revealed the relevant mechanisms. In fibroblasts, microtubule domains in the bleached zone are replaced microtubule by microtubule and not by mechanisms that affect all microtubules simultaneously. Of the models we consider, treadmilling and subunit exchange along the length do not account for this observation, but dynamic instability can since it suggests that growing and shrinking microtubules coexist. In addition, we show that the half-time for microtubule replacement is shortest at the leading edge. Dynamic instability accounts for this observation if in general microtubules do not catastrophically disassemble from the plus end, but instead have a significant probability of undergoing a transition to the growing phase before they depolymerize completely. This type of instability we call tempered rather than catastrophic because, through limited disassembly followed by regrowth, it will preferentially replace polymer domains at the ends of microtubules, thus accounting for the observation that the half-time of microtubule domain replacement is shorter with proximity to the leading edge.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Komarova, Y. A., Akhmanova, A. S., Kojima, S.-i., Galjart, N., Borisy, G. G. (2002). Cytoplasmic linker proteins promote microtubule rescue in vivo. JCB 159: 589-599 [Abstract] [Full Text]
Komarova, Y. A., Vorobjev, I. A., Borisy, G. G. (2002). Life cycle of MTs: persistent growth in the cell interior, asymmetric transition frequencies and effects of the cell boundary. J. Cell Sci. 115: 3527-3539 [Abstract] [Full Text]
Yvon, A.-M. C., Wadsworth, P. (2000). Region-Specific Microtubule Transport in Motile Cells. JCB 151: 1003-1012 [Abstract] [Full Text]
Vorobjev, I., Rodionov, V., Maly, I., Borisy, G. (1999). Contribution of plus and minus end pathways to microtubule turnover. J. Cell Sci. 112: 2277-2289 [Abstract]
Saoudi, Y., Fotedar, R., Abrieu, A., Doree, M., Wehland, J., Margolis, R. L., Job, D. (1998). Stepwise Reconstitution of Interphase Microtubule Dynamics in Permeabilized Cells and Comparison to Dynamic Mechanisms in Intact Cells. JCB 142: 1519-1532 [Abstract] [Full Text]
Blocker, A, Griffiths, G, Olivo, J., Hyman, A., Severin, F. (1998). A role for microtubule dynamics in phagosome movement. J. Cell Sci. 111: 303-312 [Abstract]
Waterman-Storer, C. M., Salmon, E.D. (1997). 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. JCB 139: 417-434 [Abstract] [Full Text]
Keating, T. J., Peloquin, J. G., Rodionov, V. I., Momcilovic, D., Borisy, G. G. (1997). Microtubule release from the�centrosome. Proc. Natl. Acad. Sci. USA 94: 5078-5083 [Abstract] [Full Text]
Rodionov, V. I., Borisy, G. G. (1997). Microtubule Treadmilling in Vivo. Science 275: 215-218 [Abstract] [Full Text]
Paramio, J., Casanova, M., Alonso, A, Jorcano, J. (1997). Keratin intermediate filament dynamics in cell heterokaryons reveals diverse behaviour of different keratins. J. Cell Sci. 110: 1099-1111 [Abstract]
Hush, J., Wadsworth, P, Callaham, D., Hepler, P. (1994). Quantification of microtubule dynamics in living plant cells using fluorescence redistribution after photobleaching. J. Cell Sci. 107: 775-784 [Abstract]
Zhai, Y, Borisy, G. (1994). Quantitative determination of the proportion of microtubule polymer present during the mitosis-interphase transition. J. Cell Sci. 107: 881-890 [Abstract]
Oakley, C, Brunette, D. (1993). The sequence of alignment of microtubules, focal contacts and actin filaments in fibroblasts spreading on smooth and grooved titanium substrata. J. Cell Sci. 106: 343-354 [Abstract]
Keates, R., Hallett, F. (1988). Dynamic instability of sheared microtubules observed by quasi-elastic light scattering. Science 241: 1642-1645 [Abstract]
Yvon, A.-M. C., Gross, D. J., Wadsworth, P. (2001). Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells. Proc. Natl. Acad. Sci. USA 98: 8656-8661 [Abstract] [Full Text]