The cortical microfilament system of lymphoblasts displays a periodic oscillatory activity in the absence of microtubules: implications for cell polarity.
For an understanding of the role of microtubules in the definition of cell polarity, we have studied the cell surface motility of human lymphoblasts (KE37 cell line) using video microscopy, time-lapse photography, and immunofluorescent localization of F-actin and myosin. Polarized cell surface motility occurs in association with a constriction ring which forms on the centrosome side of the cell: the cytoplasm flows from the ring zone towards membrane veils which keep protruding in the same general direction. This association is ensured by microtubules: in their absence the ring is conspicuous and moves periodically back and forth across the cell, while a protrusion of membrane occurs alternately at each end of the cell when the ring is at the other. This oscillatory activity is correlated with a striking redistribution of myosin towards a cortical localization and appears to be due to the alternate flow of cortical myosin associated with the ring and to the periodic assembly of actin coupled with membrane protrusion. The ring cycle involves the progressive recruitment of myosin from a polar accumulation, or cap, its transportation across the cell and its accumulation in a new cap at the other end of the cell, suggesting an assembly-disassembly process. Inhibition of actin assembly induces, on the other hand, a dramatic microtubule-dependent cell elongation with definite polarity, likely to involve the interaction of microtubules with the cell cortex. We conclude that the polarized cell surface motility in KE37 cells is based on the periodic oscillatory activity of the actin system: a myosin-powered equatorial contraction and an actin-based membrane protrusion are concerted at the cell level and occur at opposite ends of the cell in absence of microtubules. This defines a polarity which reverses periodically as the ring moves across the cell. Microtubules impose a stable cell polarity by suppressing the ring movement. A permanent association of the myosin-powered contraction and the membrane protrusion is established which results in the unidirectional activity of the actin system. Microtubules exert their effect by controlling the recruitment of cytoplasmic myosin into the cortex, probably through their direct interaction with the cortical microfilament system.
Bornens
M
M
Centre de Genetique Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Paintrand
M
M
Celati
C
C
D008841:Actin Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D000199:Actins / Q000032:analysis; D000470:Alkaloids / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D003599:Cytoskeleton / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D008961:Models, Structural; D009218:Myosins / Q000032:analysis; D015739:Nocodazole; D017239:Paclitaxel; D013601:T-Lymphocytes / Q000502:physiology / Q000648:ultrastructure
D000199:Actins; D000470:Alkaloids; D001562:Benzimidazoles; D009218:Myosins; D017239:Paclitaxel; D015739:Nocodazole