Selective stabilization of microtubules oriented toward the direction of cell migration.
A small subset of the microtubule (MT) array in many cultured cells does not exhibit the rapid turnover (t 1/2 approximately equal to 10 min) shown by most cellular MTs. The function of the stable class of MTs is unknown and has been confounded by the apparent lack of organization of stable MTs within cells. Using an antibody against detyrosinated tubulin, a post-translationally modified form of tubulin that accumulates in stable MTs, we localized the stable MTs in mouse 3T3 cells induced to initiate directional migration by experimental wounding of confluent monolayers. Immediately after monolayer wounding, the distribution of stable MTs in cells at the wound edge resembled that in cells in the monolayer interior; most cells either contained randomly distributed stable MTs or lacked them entirely. However, by 20 min after wounding, cells at the wound margin began to generate an asymmetric MT array, with virtually all stable MTs oriented toward the cell edge in contact with the wound. Two hours after monolayer wounding, greater than or equal to 80% of cells at the wound margin had generated this polarized array of stable MTs, and the array was maintained for at least 12 hr. MTs in the polarized array showed enhanced resistance to depolymerization by nocodazole, thus providing an independent test of their stability. Formation of the polar array of stable MTs appeared to precede onset of cell migration and closely paralleled reorientation of the MT-organizing center. These results show that cultured cells can remodel their MT array rapidly in response to an extracellular signal and suggest that selective stabilization of MTs is an early event in the generation of cellular asymmetry.
Gundersen
G G
GG
Department of Biology, University of California, Los Angeles 90024.
Bulinski
J C
JC
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D002465:Cell Movement; D002478:Cells, Cultured; D051379:Mice; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D015739:Nocodazole; D014404:Tubulin / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D001562:Benzimidazoles; D014404:Tubulin; D014443:Tyrosine; D015739:Nocodazole