The role of tubulin polymerization during spindle elongation in vitro

Masuda, Hisashi; Cande, W. Zacheus

doi:10.1016/0092-8674(87)90560-5

Cited by 59 publications

(65 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In previous studies we have made use of the exquisite architecture of the diatom central spindle to demonstrate that the primary mechanochemical event involved in diatom spindle elongation is the sliding apart of the MTs of the two half-spindles due to forces generated within the zone of MT overlap (3)(4)(5)10). Similar experiments using cells of permeabilized fission yeast demonstrated that a sliding mechanism may be responsible for spindle elongation in this organism as well (6).…”

Section: Discussionmentioning

confidence: 83%

“…Using functional spindles prepared from diatoms we have demonstrated that forces sufficient to drive half-spindles apart during spindle elongation (anaphase B) must be generated in the zone of antiparallel microtubule overlap (3)(4)(5). Similar forces generated by mechanochemical interactions in the zone of microtubule overlap may play a role during mitosis in most eukaryotic cells (6)(7)(8)(9).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain.

Hogan

Wein

Wordeman

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Isolated central spindles or spindles in detergent-permeabilized celis from the diatom Cylindrotheca fusifornis can undergo ATP-dependent reactivation of spindle elongation in vitro. We have used a peptide antibody raised agains a 10-amino acid portion common to the kinesin superfamily motor domain to look for kinesin-like motor activity during anaphase B of mitosis. The peptide antibody localizes to central spindles. Upon ATP reactivation of spindle elongation, antigens recognized by the antibody are associated exclusively with the central spindle midzone where antiparaHlel microtubuies of each half-spindle overlap. The antibody recognizes several polypeptides by immunoblot using isolated spindle extracts. One of these polypeptides behaves like kinesin with respect to nucleotide-specific binding to and release from taxol-stabilized microtubules. Preincubation of the spindle model with the peptide antibody inhibits subsequent ATP reactivation of spindle elongation. Coincubation of the peptide antibody with peptide antigen rescues spindle function. These results support a role for kinesin-related protein(s) in spindle elongation (anaphase B) of mitosis and suggest that one or several polypeptides that we have identified in spindle extracts may fulfill this function.Diatom spindles are important model systems for describing the rearrangements of microtubules (MTs) that occur during anaphase because the MTs associated with spindle elongation are closely packed in an antiparallel overlapping MT array (the central spindle) and are spatially separated from kinetochore and astral MTs (1, 2). Using functional spindles prepared from diatoms we have demonstrated that forces sufficient to drive half-spindles apart during spindle elongation (anaphase B) must be generated in the zone of antiparallel microtubule overlap (3-5). Similar forces generated by mechanochemical interactions in the zone of microtubule overlap may play a role during mitosis in most eukaryotic cells (6-9). The pennate diatom Cylindrothecafusiformis can be readily grown in bulk cultures suitable for biochemistry and the cells can be arrested in mitosis by a combination of light/dark cell cycle synchronization and drug arrest. At harvest, 40-60% of the cells contain 4-to 5-,Am-long metaphase/early anaphase spindles and interphase MT arrays are absent (10). After isolation or cell permeabilization the two overlapping half-spindles that comprise the central spindle retain the ability to slide apart when incubated with 1 mM

show abstract

Section: Discussionmentioning

confidence: 83%

mentioning

confidence: 99%

Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain.

Hogan

Wein

Wordeman

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Because severed minus-ends of spindle microtubules are stable in grasshopper spermatocytes (Nicklas, 1989), we think that new half-spindle elongation occurs because of the growth of plus-ends of microtubules. Following its establishment, addition of tubulin at plus-ends along with motor-driven antagonistic sliding of microtubules (Masuda and Cande, 1987;Cande and Hogan, 1989;Nislow et al, 1992;Sawin and Endow, 1993;Sharp et al, 1999;Mishima et al, 2002;Dechant and Glotzer, 2003;Goshima and Vale, 2003) may extend the new half-spindle away from microtubule overlap at the midzone. This apparent microtubule extrusion process may occur much as described for formation of overlapping central spindle microtubules from chromatin beads in Xenopus egg extracts (Heald et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Microtubules continuously dictate distribution of actin filaments and positioning of cell cleavage in grasshopper spermatocytes

Alsop

Zhang

2004

Journal of Cell Science

View full text Add to dashboard Cite

We systematically examined the impact of microtubules on distribution of actin filaments and positioning of cell cleavage using micromanipulation to progressively alter the symmetric distribution of spindle microtubules in grasshopper spermatocytes. The initial microtubule asymmetry was induced by placing a single chromosome at one spindle pole using a microneedle, which facilitates regional assembly of spindle microtubules. We augmented chromosome-induced microtubule asymmetry by further removing the aster from the achromosomal pole, producing unichromosome-bearing monopolar spindles. We created the highest spindle asymmetry by cutting early anaphase cells in two, each containing a full set of segregating chromosomes in a half-spindle. We demonstrate that the location of the spindle midzone, distribution of actin filaments, and position of cell cleavage depend on the amount of microtubule asymmetry generated, shifting up to 48.6±3.8% away from the spindle equator in cut cells. The positional shift is dynamic, changing incessantly as spindle microtubules reorganize during cytokinesis. These results suggest that microtubules continuously dictate the distribution of actin filaments and positioning of cell cleavage in grasshopper spermatocytes.

show abstract

“…This finding indicates that the regulation of microtubule dynamics is important in determining the rate of spindle elongation. Previous evidence suggests that microtubule growth rate factors can control the rate of spindle elongation (Masuda and Cande, 1987). Experiments in yeast have shown that the biphasic nature of spindle elongation results from the balanced influence of various microtubule motor proteins (Straight et al, 1998).…”

Section: Spindle Dynamics In Tub2-c354 Mutant Cellsmentioning

confidence: 99%

β-Tubulin C354 Mutations that Severely Decrease Microtubule Dynamics Do Not Prevent Nuclear Migration in Yeast

Gupta

Bode

Thrower

et al. 2002

MBoC

View full text Add to dashboard Cite

Microtubule dynamics are influenced by interactions of microtubules with cellular factors and by changes in the primary sequence of the tubulin molecule. Mutations of yeast ␤-tubulin C354, which is located near the binding site of some antimitotic compounds, reduce microtubule dynamicity greater than 90% in vivo and in vitro. The resulting intrinsically stable microtubules allowed us to determine which, if any, cellular processes are dependent on dynamic microtubules. The average number of cytoplasmic microtubules decreased from 3 in wild-type to 1 in mutant cells. The single microtubule effectively located the bud site before bud emergence. Although spindles were positioned near the bud neck at the onset of anaphase, the mutant cells were deficient in preanaphase spindle alignment along the mother-bud axis. Spindle microtubule dynamics and spindle elongation rates were also severely depressed in the mutants. The pattern and extent of cytoplasmic microtubule dynamics modulation through the cell cycle may reveal the minimum dynamic properties required to support growth. The ability to alter intrinsic microtubule dynamics and determine the in vivo phenotype of cells expressing the mutant tubulin provides a critical advance in assessing the dynamic requirements of an essential gene function.

show abstract

The role of tubulin polymerization during spindle elongation in vitro

Cited by 59 publications

References 34 publications

Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain.

Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain.

Microtubules continuously dictate distribution of actin filaments and positioning of cell cleavage in grasshopper spermatocytes

β-Tubulin C354 Mutations that Severely Decrease Microtubule Dynamics Do Not Prevent Nuclear Migration in Yeast

Contact Info

Product

Resources

About