Microtubule-associated motility in cytoplasmic extracts of sea urchin eggs

Cell Motility and the Cytoskeleton
N R Gliksman, E D Salmon

Abstract

We have developed a method for producing sea urchin egg cytoplasmic extracts which support substantial microtubule-associated motility, particularly minus end-directed motility characteristic of cytoplasmic dynein. Particles translocated along microtubules and axonemes predominantly in the minus end direction; microtubules and axonemes glided across the coverslip surface only in the plus end direction (as expected for a minus-end directed motor bound to the coverslip surface); and microtubules crosslinked into bundles in an antiparallel orientation. Velocities of particle and microtubule translocation were in the range of 0.5-1.8 microns/sec. Vanadate at 10 microM inhibited all gliding of the microtubules and axonemes, yet bidirectional particle transport persisted. Vanadate at concentrations of 25 microM and higher inhibited nearly all microtubule-based motility in the preparation and produced parallel bundling of the microtubules. Motility was slowed but not stopped in the presence of 5 mM AMP-PNP. Usually when a particle bound to a microtubule wall, it moved to the microtubule minus end. These particles often remained attached to the minus end. When a microtubule plus end in the shortening phase of dynamic instability reache...Continue Reading

References

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Citations

Jan 1, 1996·Cell Motility and the Cytoskeleton·G R Rosania, J A Swanson
Jan 1, 1996·Cell Motility and the Cytoskeleton·C S Spittle, L Cassimeris
Jan 1, 1995·The Journal of Cell Biology·A Desai, T J Mitchison
Dec 1, 1995·Molecular Biology of the Cell·S Inoué, E D Salmon
Jul 9, 2011·Molecular Biology of the Cell·Aaron C GroenTimothy J Mitchison

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