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Actin-dependent lamellipodia formation and microtubule-dependent tail retraction control-directed cell migration

Ballestrem, Christoph
Published in Molecular Biology of the Cell. 2000, vol. 11, no. 9, p. 2999-3012
Abstract Migrating cells are polarized with a protrusive lamella at the cell front followed by the main cell body and a retractable tail at the rear of the cell. The lamella terminates in ruffling lamellipodia that face the direction of migration. Although the role of actin in the formation of lamellipodia is well established, it remains unclear to what degree microtubules contribute to this process. Herein, we have studied the contribution of microtubules to cell motility by time-lapse video microscopy on green flourescence protein-actin- and tubulin-green fluorescence protein-transfected melanoma cells. Treatment of cells with either the microtubule-disrupting agent nocodazole or with the stabilizing agent taxol showed decreased ruffling and lamellipodium formation. However, this was not due to an intrinsic inability to form ruffles and lamellipodia because both were restored by stimulation of cells with phorbol 12-myristate 13-acetate in a Rac-dependent manner, and by stem cell factor in melanoblasts expressing the receptor tyrosine kinase c-kit. Although ruffling and lamellipodia were formed without microtubules, the microtubular network was needed for advancement of the cell body and the subsequent retraction of the tail. In conclusion, we demonstrate that the formation of lamellipodia can occur via actin polymerization independently of microtubules, but that microtubules are required for cell migration, tail retraction, and modulation of cell adhesion.
Keywords Actins/genetics/ physiologyAnimalsCell LineCell Movement/drug effects/ physiologyCytoplasm/physiology/ultrastructureGreen Fluorescent ProteinsLuminescent Proteins/analysis/geneticsMelanocytesMelanoma, ExperimentalMiceMicrotubules/drug effects/ physiology/ultrastructureNocodazole/pharmacologyPaclitaxel/pharmacologyProto-Oncogene Proteins c-kit/metabolismRecombinant Fusion Proteins/analysis/biosynthesisTetradecanoylphorbol Acetate/pharmacologyTransfectionTumor Cells, Culturedrac GTP-Binding Proteins/metabolism
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PMID: 10982396
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