Interaction of vinblastine with steady-state microtubules in vitro

Wilson, Leslie; Jordan, Mary Ann; Morse, Aileen N. C.; Margolis, Robert L.

doi:10.1016/0022-2836(82)90035-3

Cited by 114 publications

(73 citation statements)

References 32 publications

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“…Vinblastine at low concentrations stabilizes plus ends but not minus ends. The maximum number of high affinity vinblastine sites at microtubule ends determined by extrapolation of binding data to infinitely high vinblastine concentration is small (16 sites per microtubule (25)); this value is within experimental error of the protofilament number in reconstituted microtubules in vitro (approximately 14 -15 protofilaments (42)). In addition, the tubulin dimer has structural polarity and the microtubule ends differ kinetically.…”

Section: Fig 3 Microtubule Length Changes Per Growing (A) or Shortementioning

confidence: 82%

“…How Does Vinblastine Stabilize Microtubule Dynamics at Plus Ends?-It is known from previous studies that at low concentrations vinblastine binds reversibly and with relatively high affinity directly to at least one and perhaps both microtubule ends, without being incorporated into the core of the polymer (25). Binding of vinblastine to tubulin is also known to induce conformational changes in tubulin (34 -37).…”

Section: Fig 3 Microtubule Length Changes Per Growing (A) or Shortementioning

confidence: 99%

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Differential Effects of Vinblastine on Polymerization and Dynamics at Opposite Microtubule Ends

Panda

Jordan

Chu

et al. 1996

Journal of Biological Chemistry

117

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We have characterized the effects of vinblastine on the growing and shortening dynamics at opposite ends of individual bovine brain microtubules at steady state in vitro by video microscopy. Vinblastine exerted strikingly different effects on the dynamics and polymer mass at the plus and minus ends of microtubules. At concentrations between 0.1 and 0.4 M, the drug strongly depolymerized microtubules at minus ends, whereas it did not significantly depolymerize microtubules at plus ends. Vinblastine stabilized plus ends by suppressing the rate and extent of growth and shortening, decreasing the catastrophe frequency, and increasing the rescue frequency. In contrast, vinblastine destabilized minus ends by increasing the catastrophe frequency and decreasing the rescue frequency, whereas it had no effect on the rate or extent of growth or shortening. Thus, vinblastine moderately increased the overall dynamicity at minus ends while strongly suppressing dynamicity at plus ends. Both the kinetic destabilization of microtubules at minus ends and the stabilization at plus ends may contribute to the altered function of mitotic spindle microtubules of cells blocked in mitosis by low concentrations of vinblastine.

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Section: Fig 3 Microtubule Length Changes Per Growing (A) or Shortementioning

confidence: 82%

Section: Fig 3 Microtubule Length Changes Per Growing (A) or Shortementioning

confidence: 99%

Differential Effects of Vinblastine on Polymerization and Dynamics at Opposite Microtubule Ends

Panda

Jordan

Chu

et al. 1996

Journal of Biological Chemistry

117

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“…The one exception was seen at the plus ends, after treatment with 100 nM nocodazole; rescue increased slightly in this case. (Wilson et al, 1982;Jordan et al, 1991;Toso et al, 1993), nocodazole binds to MTs and kinetically caps the MT end, then there should be an increase in resistance to Figure 5). In fact, nearly all MTs had disappeared by the time focus was restored.…”

Section: Nocodazole Suppresses Mt Dynamic Instability In Interphase Cmentioning

confidence: 98%

“…This mechanism is different from that proposed for vinblastine. The effects of low concentrations of vinblastine on MT dynamics are believed to result from the binding of vinblastine to high-affinity sites at or near the growing end of the MT (Wilson et al, 1982;Jordan and Wilson, 1990;Toso et al, 1993; and not on binding to the tubulin dimer. The mechanism of action of nocodazole may be more similar to colchicine whose effects on MT assembly are mediated by a tubulin-colchicine complex (Wilson and Farrell, 1986;Skoufias et al, 1992).…”

Section: Nanomolar Concentrations Of Nocodazole Influencementioning

confidence: 99%

Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro.

Vasquez

Howell

Yvon

et al. 1997

MBoC

408

328

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Previous studies demonstrated that nanomolar concentrations of nocodazole can block cells in mitosis without net microtubule disassembly and resulted in the hypothesis that this block was due to a nocodazole-induced stabilization of microtubules. We tested this hypothesis by examining the effects of nanomolar concentrations of nocodazole on microtubule dynamic instability in interphase cells and in vitro with purified brain tubulin. Newt lung epithelial cell microtubules were visualized by video-enhanced differential interference contrast microscopy and cells were perfused with solutions of nocodazole ranging in concentration from 4 to 400 nM. Microtubules showed a loss of the two-state behavior typical of dynamic instability as evidenced by the addition of a third state where they exhibited little net change in length (a paused state). Nocodazole perfusion also resulted in slower elongation and shortening velocities, increased catastrophe, and an overall decrease in microtubule turnover. Experiments performed on BSC-1 cells that were microinjected with rhodamine-labeled tubulin, incubated in nocodazole for 1 h, and visualized by using low-light-level fluorescence microscopy showed similar results except that nocodazole-treated BSC-1 cells showed a decrease in catastrophe. To gain insight into possible mechanisms responsible for changes in dynamic instability, we examined the effects of 4 nM to 12 ,uM nocodazole on the assembly of purified tubulin from axoneme seeds. At both microtubule plus and minus ends, perfusion with nocodazole resulted in a dose-dependent decrease in elongation and shortening velocities, increase in pause duration and catastrophe frequency, and decrease in rescue frequency. These effects, which result in an overall decrease in microtubule turnover after nocodazole treatment, suggest that the mitotic block observed is due to a reduction in microtubule dynamic turnover. In addition, the in vitro results are similar to the effects of increasing concentrations of GDP-tubulin (TuD) subunits on microtubule assembly. Given that nocodazole increases tubulin GTPase activity, we propose that nocodazole acts by generating TuD subunits that then alter dynamic instability.

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“…TX binds and stabilizes microtubule polymers, thereby preventing microtubule dissociation (36). VB is a plant alkaloid that binds to tubulin and prevents microtubule assembly at concentrations at or below 10 M (21,24,34,44). CN binds to tubulin and blocks microtubule formation by stimulating the intrinsic GTPase activity of tubulin (5,25).…”

mentioning

confidence: 99%

Effects of Microtubule Modulators on HIV-1 Infection of Transformed and Resting CD4 T Cells

Yoder

Guo

et al. 2011

J Virol

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Interaction of vinblastine with steady-state microtubules in vitro

Cited by 114 publications

References 32 publications

Differential Effects of Vinblastine on Polymerization and Dynamics at Opposite Microtubule Ends

Differential Effects of Vinblastine on Polymerization and Dynamics at Opposite Microtubule Ends

Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro.

Effects of Microtubule Modulators on HIV-1 Infection of Transformed and Resting CD4 T Cells

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