scholarly journals Okadaic acid induces interphase to mitotic-like microtubule dynamic instability by inactivating rescue.

1992 ◽  
Vol 119 (5) ◽  
pp. 1271-1276 ◽  
Author(s):  
N R Gliksman ◽  
S F Parsons ◽  
E D Salmon
Keyword(s):  
High Resolution ◽  
Sea Urchin ◽  
Okadaic Acid ◽  
Dynamic Instability ◽  
Major Change ◽  
Microtubule Dynamics ◽  
Regulatory Pathway ◽  
Microtubule Dynamic ◽  
Interphase Cells ◽  
Frequent Switching

We used high-resolution video microscopy to visualize microtubule dynamic instability in extracts of interphase sea urchin eggs and to analyze the changes that occur upon addition of 0.8-2.5 microM okadaic acid, an inhibitor of phosphatase 1 and 2A (PP1, PP2a) (Bialojan, D., and A. Takai. 1988. Biochem. J. 256:283-290). Microtubule plus-ends in these extracts oscillated between the elongation and shortening phases of dynamic instability at frequencies typical for interphase cells. Switching from elongation to shortening (catastrophe) was frequent, but microtubules persisted and grew long because of frequent switching back to elongation (rescue). Addition of okadaic acid to the extract induced rapid (< 5 min) conversion to short, dynamic microtubules typical of mitosis. The frequency of catastrophe doubled and the velocities of elongation and shortening increased slightly; however, the major change was an elimination of rescue. Thus, modulation of the rescue frequency by phosphorylation-dependent mechanisms may be a major regulatory pathway for selectively controlling microtubule dynamics without dramatically changing velocities of microtubule elongation and shortening.

scholarly journals Predicted effects of severing enzymes on the length distribution and total mass of microtubules

2019 ◽  
Author(s):  
Y.-W. Kuo ◽  
O. Trottier ◽  
J. Howard
Keyword(s):  
Steady State ◽  
Numerical Solutions ◽  
Dynamic Instability ◽  
Computational Simulation ◽  
Length Distribution ◽  
Microtubule Dynamics ◽  
Microtubule Dynamic ◽  
Cytoskeletal Network ◽  
Hydrolyzing Enzymes ◽  
In Cells

AbstractMicrotubules are dynamic cytoskeletal polymers whose growth and shrinkage are highly regulated as eukaryotic cells change shape, move and divide. One family of microtubule regulators includes the ATP-hydrolyzing enzymes spastin, katanin and fidgetin, which sever microtubule polymers into shorter fragments. Paradoxically, severases can increase microtubule number and mass in cells. Recent work with purified spastin and katanin accounts for this phenotype by showing that, in addition to severing, these enzymes modulate microtubule dynamics by accelerating the conversion of microtubules to the growing state and thereby promoting their regrowth. This leads to the observed exponential increase in microtubule mass. Spastin also influences the steady-state distribution of microtubule lengths, changing it from an exponential, as predicted by models of microtubule dynamic instability, to a peaked distribution. This effect of severing and regrowth by spastin on the microtubule length distribution has not been explained theoretically. To solve this problem, we formulated and solved a master equation for the time evolution of microtubule lengths in the presence of severing and microtubule dynamic instability. We then obtained numerical solutions to the steady-state length distribution and showed that the rate of severing and the speed of microtubule growth are the dominant parameters determining the steady-state length distribution. Furthermore, we found that the amplification rate is predicted to increase with severing, which is a new result. Our results establish a theoretical basis for how severing and dynamics together can serve to nucleate new microtubules, constituting a versatile mechanism to regulate microtubule length and mass.SignificanceThe numbers and lengths of microtubules are tightly regulated in cells. Severing enzymes fragment microtubules into shorter filaments and are important for cell division and tissue development. Previous work has shown that severing can lead to an increase in total microtubule number and mass, but the effect of severing on microtubule length is not understood quantitatively. Combining mathematical modeling and computational simulation, we solve the microtubule length distribution in the presence of severing enzymes and explore how severing activity and microtubule dynamics collectively control microtubule number and length. These results advance our understanding of the physical basis of severing as a regulatory mechanism shaping the cellular cytoskeletal network.

scholarly journals Asymmetric behavior of severed microtubule ends after ultraviolet-microbeam irradiation of individual microtubules in vitro.

1989 ◽  
Vol 108 (3) ◽  
pp. 931-937 ◽  
Author(s):  
R A Walker ◽  
S Inoué ◽  
E D Salmon
Keyword(s):  
Sea Urchin ◽  
Molecular Basis ◽  
Dynamic Instability ◽  
Second Step ◽  
Microtubule Dynamic ◽  
Asymmetric Behavior ◽  
Ultraviolet Microbeam ◽  
Cap Model ◽  
Flagellar Axoneme

The molecular basis of microtubule dynamic instability is controversial, but is thought to be related to a "GTP cap." A key prediction of the GTP cap model is that the proposed labile GDP-tubulin core will rapidly dissociate if the GTP-tubulin cap is lost. We have tested this prediction by using a UV microbeam to cut the ends from elongating microtubules. Phosphocellulose-purified tubulin was assembled onto the plus and minus ends of sea urchin flagellar axoneme fragments at 21-22 degrees C. The assembly dynamics of individual microtubules were recorded in real time using video microscopy. When the tip of an elongating plus end microtubule was cut off, the severed plus end microtubule always rapidly shortened back to the axoneme at the normal plus end rate. However, when the distal tip of an elongating minus end microtubule was cut off, no rapid shortening occurred. Instead, the severed minus end resumed elongation at the normal minus end rate. Our results show that some form of "stabilizing cap," possibly a GTP cap, governs the transition (catastrophe) from elongation to rapid shortening at the plus end. At the minus end, a simple GTP cap is not sufficient to explain the observed behavior unless UV induces immediate recapping of minus, but not plus, ends. Another possibility is that a second step, perhaps a structural transformation, is required in addition to GTP cap loss for rapid shortening to occur. This transformation would be favored at plus, but not minus ends, to account for the asymmetric behavior of the ends.

scholarly journals A mutation uncouples the tubulin conformational and GTPase cycles, revealing allosteric control of microtubule dynamics

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Elisabeth A Geyer ◽  
Alexander Burns ◽  
Beth A Lalonde ◽  
Xuecheng Ye ◽  
Felipe-Andres Piedra ◽  
...  
Keyword(s):  
Conformational Change ◽  
Dynamic Instability ◽  
Dynamic Properties ◽  
Microtubule Dynamics ◽  
Gtpase Activity ◽  
Microtubule Dynamic ◽  
Gtp Hydrolysis ◽  
Allosteric Control

Microtubule dynamic instability depends on the GTPase activity of the polymerizing αβ-tubulin subunits, which cycle through at least three distinct conformations as they move into and out of microtubules. How this conformational cycle contributes to microtubule growing, shrinking, and switching remains unknown. Here, we report that a buried mutation in αβ-tubulin yields microtubules with dramatically reduced shrinking rate and catastrophe frequency. The mutation causes these effects by suppressing a conformational change that normally occurs in response to GTP hydrolysis in the lattice, without detectably changing the conformation of unpolymerized αβ-tubulin. Thus, the mutation weakens the coupling between the conformational and GTPase cycles of αβ-tubulin. By showing that the mutation predominantly affects post-GTPase conformational and dynamic properties of microtubules, our data reveal that the strength of the allosteric response to GDP in the lattice dictates the frequency of catastrophe and the severity of rapid shrinking.

scholarly journals Vinblastine suppresses dynamics of individual microtubules in living interphase cells.

1995 ◽  
Vol 6 (9) ◽  
pp. 1215-1229 ◽  
Author(s):  
R Dhamodharan ◽  
M A Jordan ◽  
D Thrower ◽  
L Wilson ◽  
P Wadsworth
Keyword(s):  
Dynamic Instability ◽  
Living Cells ◽  
Microtubule Dynamics ◽  
Microtubule Depolymerization ◽  
Dynamic Activity ◽  
Low Concentrations ◽  
Interphase Cells ◽  
Endogenous Regulators ◽  
Microtubule Growth

We have characterized the effects of vinblastine on the dynamic instability behavior of individual microtubules in living BS-C-1 cells microinjected with rhodamine-labeled tubulin and have found that at low concentrations (3-64 nM), vinblastine potently suppresses dynamic instability without causing net microtubule depolymerization. Vinblastine suppressed the rates of microtubule growth and shortening, and decreased the frequency of transitions from growth or pause to shortening, also called catastrophe. In vinblastine-treated cells, both the average duration of a pause (a state of attenuated dynamics where neither growth nor shortening could be detected) and the percentage of total time spent in pause were significantly increased. Vinblastine potently decreased dynamicity, a measure of the overall dynamic activity of microtubules, reducing this parameter by 75% at 32 nM. The present work, consistent with earlier in vitro studies, demonstrates that vinblastine kinetically caps the ends of microtubules in living cells and supports the hypothesis that the potent chemotherapeutic action of vinblastine as an antitumor drug is suppression of mitotic spindle microtubule dynamics. Further, the results indicate that molecules that bind to microtubule ends can regulate microtubule dynamic behavior in living cells and suggest that endogenous regulators of microtubule dynamics that work by similar mechanisms may exist in living cells.

Analysis of the Mechanism of Microtubule Dynamic Instability Using a UV Microbeam to Sever Elongating Microtubules

1988 ◽  
Vol 46 ◽  
pp. 122-123
Author(s):  
R.A Walker ◽  
S. Inoue ◽  
E.D. Salmon
Keyword(s):  
Dynamic Instability ◽  
Microtubule Dynamic ◽  
Gtp Hydrolysis ◽  
Abrupt Transition ◽  
Microtubule Associated Proteins ◽  
Asymmetrical Structure ◽  
Associated Proteins

Microtubules polymerized in vitro from tubulin purified free of microtubule-associated proteins exhibit dynamic instability (1,2,3). Free microtubule ends exist in persistent phases of elongation or rapid shortening with infrequent, but, abrupt transitions between these phases. The abrupt transition from elongation to rapid shortening is termed catastrophe and the abrupt transition from rapid shortening to elongation is termed rescue. A microtubule is an asymmetrical structure. The plus end grows faster than the minus end. The frequency of catastrophe of the plus end is somewhat greater than the minus end, while the frequency of rescue of the plus end in much lower than for the minus end (4).The mechanism of catastrophe is controversial, but for both the plus and minus microtubule ends, catastrophe is thought to be dependent on GTP hydrolysis. Microtubule elongation occurs by the association of tubulin-GTP subunits to the growing end. Sometime after incorporation into an elongating microtubule end, the GTP is hydrolyzed to GDP, yielding a core of tubulin-GDP capped by tubulin-GTP (“GTP-cap”).

scholarly journals Temperature-jump studies of microtubule dynamic instability.

1988 ◽  
Vol 263 (21) ◽  
pp. 10344-10352
Author(s):  
M Caplow ◽  
J Shanks ◽  
R L Ruhlen
Keyword(s):  
Temperature Jump ◽  
Dynamic Instability ◽  
Microtubule Dynamic

scholarly journals Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle

1994 ◽  
Vol 124 (3) ◽  
pp. 223-233 ◽  
Author(s):  
CL Rieder ◽  
ED Salmon
Keyword(s):  
Mitotic Spindle ◽  
Constant Velocity ◽  
Molecular Mechanisms ◽  
Dynamic Instability ◽  
Microtubule Dynamic ◽  
Pole Motion ◽  
Microtubule Motors ◽  
Chromosome Position ◽  
Pulling Forces

We argue that hypotheses for how chromosomes achieve a metaphase alignment, that are based solely on a tug-of-war between poleward pulling forces produced along the length of opposing kinetochore fibers, are no longer tenable for vertebrates. Instead, kinetochores move themselves and their attached chromosomes, poleward and away from the pole, on the ends of relatively stationary but shortening/elongating kinetochore fiber microtubules. Kinetochores are also "smart" in that they switch between persistent constant-velocity phases of poleward and away from the pole motion, both autonomously and in response to information within the spindle. Several molecular mechanisms may contribute to this directional instability including kinetochore-associated microtubule motors and kinetochore microtubule dynamic instability. The control of kinetochore directional instability, to allow for congression and anaphase, is likely mediated by a vectorial mechanism whose magnitude and orientation depend on the density and orientation or growth of polar microtubules. Polar microtubule arrays have been shown to resist chromosome poleward motion and to push chromosomes away from the pole. These "polar ejection forces" appear to play a key role in regulating kinetochore directional instability, and hence, positions achieved by chromosomes on the spindle.

Bathymetry and uplift rate of the Gulf of Aqaba, Dead Sea Fault.

2021 ◽  
Author(s):  
Matthieu Ribot ◽  
Yann Klinger ◽  
Edwige Pons-Branchu ◽  
Marthe Lefevre ◽  
Sigurjón Jónsson
Keyword(s):  
High Resolution ◽  
Tectonic Evolution ◽  
Active Faults ◽  
Major Change ◽  
Dead Sea ◽  
Fault System ◽  
Arabian Plate ◽  
Gulf Of Aqaba ◽  
Uplift Rate ◽  
The Dead

<p>Initially described in the late 50’s, the Dead Sea Fault system connects at its southern end to the Red Sea extensive system, through a succession of left-stepping faults. In this region, the left-lateral differential displacement of the Arabian plate with respect to the Sinai micro-plate along the Dead Sea fault results in the formation of a depression corresponding to the Gulf Aqaba. We acquired new bathymetric data in the areas of the Gulf of Aqaba and Strait of Tiran during two marine campaigns (June 2018, September 2019) in order to investigate the location of the active faults, which structure and control the morphology of the area. The high-resolution datasets (10-m posting) allow us to present a new fault map of the gulf and to discuss the seismic potential of the main active faults.</p><p>We also investigated the eastern margin of the Gulf of Aqaba and Tiran island to assess the vertical uplift rate. To do so, we computed high-resolution topographic data and we processed new series of U-Th analyses on corals from the uplifted marine terraces.</p><p>Combining our results with previous studies, we determined the local and the regional uplift in the area of the Gulf of Aqaba and Strait of Tiran.</p><p>Eventually, we discussed the tectonic evolution of the gulf since the last major change of the tectonic regime and we propose a revised tectonic evolution model of the area.</p><p> </p>

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