Microtubule Flux in Mitosis is Independent of Chromosomes, Centrosomes, and Antiparallel Microtubules

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 1994 Feb 1

PMID 8019007

Citations 31

Authors

K E Sawin

T J Mitchison

Affiliations

Soon will be listed here.

Abstract

We investigated the mechanism of poleward microtubule flux in the mitotic spindle by generating spindle subassemblies in Xenopus egg extracts in vitro and assaying their ability to flux by photoactivation of fluorescence and low-light multichannel fluorescence video-microscopy. We find that monopolar intermediates of in vitro spindle assembly (half-spindles) exhibit normal poleward flux, as do astral microtubule arrays induced by the addition of dimethyl sulfoxide to egg extracts in the absence of both chromosomes and conventional centrosomes. Immunodepletion of the kinesin-related microtubule motor protein Eg5, a candidate flux motor, suggests that Eg5 is not required for flux. These results suggest that poleward flux is a basic element of microtubule behavior exhibited by even simple self-organized microtubule arrays and presumably underlies the most elementary levels of spindle morphogenesis.

Citing Articles

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Microtubule poleward flux in human cells is driven by the coordinated action of four kinesins.

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References

Inoue S, Sato H . Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement. J Gen Physiol. 1967; 50(6):Suppl:259-92. PMC: 2225745. View

McNally F, Vale R . Identification of katanin, an ATPase that severs and disassembles stable microtubules. Cell. 1993; 75(3):419-29. DOI: 10.1016/0092-8674(93)90377-3. View

Inoue S, RITTER Jr H . Dynamics of mitotic spindle organization and function. Soc Gen Physiol Ser. 1975; 30:3-30. View

Margolis R, Wilson L . Opposite end assembly and disassembly of microtubules at steady state in vitro. Cell. 1978; 13(1):1-8. DOI: 10.1016/0092-8674(78)90132-0. View

Margolis R, Wilson L . Microtubule treadmills--possible molecular machinery. Nature. 1981; 293(5835):705-11. DOI: 10.1038/293705a0. View

Inoue S . Cell division and the mitotic spindle. J Cell Biol. 1981; 91(3 Pt 2):131s-147s. PMC: 2112807. DOI: 10.1083/jcb.91.3.131s. View

Leslie R, Pickett-Heaps J . Spindle microtubule dynamics following ultraviolet-microbeam irradiations of mitotic diatoms. Cell. 1984; 36(3):717-27. DOI: 10.1016/0092-8674(84)90352-0. View

Salmon E, Leslie R, Saxton W, Karow M, McIntosh J . Spindle microtubule dynamics in sea urchin embryos: analysis using a fluorescein-labeled tubulin and measurements of fluorescence redistribution after laser photobleaching. J Cell Biol. 1984; 99(6):2165-74. PMC: 2113564. DOI: 10.1083/jcb.99.6.2165. View

Saxton W, Stemple D, Leslie R, Salmon E, Zavortink M, McIntosh J . Tubulin dynamics in cultured mammalian cells. J Cell Biol. 1984; 99(6):2175-86. PMC: 2113582. DOI: 10.1083/jcb.99.6.2175. View

10.

Mitchison T, Kirschner M . Dynamic instability of microtubule growth. Nature. 1984; 312(5991):237-42. DOI: 10.1038/312237a0. View

11.

Hill T . Theoretical problems related to the attachment of microtubules to kinetochores. Proc Natl Acad Sci U S A. 1985; 82(13):4404-8. PMC: 391109. DOI: 10.1073/pnas.82.13.4404. View

12.

Mitchison T, Kirschner M . Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation. J Cell Biol. 1985; 101(3):766-77. PMC: 2113737. DOI: 10.1083/jcb.101.3.766. View

13.

Rieder C, Davison E, Jensen L, Cassimeris L, Salmon E . Oscillatory movements of monooriented chromosomes and their position relative to the spindle pole result from the ejection properties of the aster and half-spindle. J Cell Biol. 1986; 103(2):581-91. PMC: 2113830. DOI: 10.1083/jcb.103.2.581. View

14.

Koshland D, Mitchison T, Kirschner M . Polewards chromosome movement driven by microtubule depolymerization in vitro. Nature. 1988; 331(6156):499-504. DOI: 10.1038/331499a0. View

15.

Tao W, Walter R, Berns M . Laser-transected microtubules exhibit individuality of regrowth, however most free new ends of the microtubules are stable. J Cell Biol. 1988; 107(3):1025-35. PMC: 2115282. DOI: 10.1083/jcb.107.3.1025. View

16.

Walker R, OBrien E, Pryer N, Soboeiro M, Voter W, Erickson H . Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies. J Cell Biol. 1988; 107(4):1437-48. PMC: 2115242. DOI: 10.1083/jcb.107.4.1437. View

17.

Hotani H, Horio T . Dynamics of microtubules visualized by darkfield microscopy: treadmilling and dynamic instability. Cell Motil Cytoskeleton. 1988; 10(1-2):229-36. DOI: 10.1002/cm.970100127. View

18.

Walker R, Inoue S, Salmon E . Asymmetric behavior of severed microtubule ends after ultraviolet-microbeam irradiation of individual microtubules in vitro. J Cell Biol. 1989; 108(3):931-7. PMC: 2115382. DOI: 10.1083/jcb.108.3.931. View

19.

Mitchison T . Polewards microtubule flux in the mitotic spindle: evidence from photoactivation of fluorescence. J Cell Biol. 1989; 109(2):637-52. PMC: 2115701. DOI: 10.1083/jcb.109.2.637. View

20.

Wilson P, Forer A . Ultraviolet microbeam irradiation of chromosomal spindle fibres shears microtubules and permits study of the new free ends in vivo. J Cell Sci. 1988; 91 ( Pt 4):455-68. DOI: 10.1242/jcs.91.4.455. View