Centrosomal control of microtubule dynamics.
In many animal cells, minus ends of microtubules (MTs) are thought to be capped by the centrosome whereas plus ends are free and display dynamic instability. We tested the role of the centrosome by examining MT behavior in cytoplasts from which the centrosome was removed. Cells were injected with Cy3-tubulin to fluorescently label MTs and were enucleated by using a centrifugation procedure. Enucleation resulted in a mixture of cytoplasts containing or lacking the centrosome. Fibroblast (CHO-K1) and epithelial (BSC-1) cells were investigated. In fibroblast cytoplasts containing the centrosome, MTs showed dynamic instability indistinguishable from that in intact cells. In contrast, in cytoplasts lacking the centrosome, MTs treadmilled-shortened at the minus end at about 12 micrometers/min while growing at the plus end at the same rate. The change in behavior of the plus end from dynamic instability to persistent growth correlated with an elevated level of free tubulin subunits (78% in centrosome-free cytoplasts vs. 44% in intact cells) generated by minus-end depolymerization. In contrast to fibroblast cells, in centrosome-free cytoplasts prepared from epithelial cells, MTs displayed dynamic instability at plus ends and relative stability at minus ends presumably because of specific minus-end stability factors distributed throughout the cytoplasm. We suggest that, in fibroblast cells, a minus-end depolymerization mechanism functions to eliminate errors in MT organization and that dynamic instability of MT plus ends is a result of capping of minus ends by the centrosome.
Rodionov
V
V
Laboratory of Molecular Biology, R. M. Bock Laboratories, University of Wisconsin, Madison, WI 53706, USA.
Nadezhdina
E
E
Borisy
G
G
D000818:Animals; D016466:CHO Cells; D002467:Cell Nucleus; D018385:Centrosome / Q000502:physiology*; D006224:Cricetinae; D003593:Cytoplasm; D004847:Epithelial Cells; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D008954:Models, Biological