Actin depolymerisation induces process formation on MAP2-transfected non-neuronal cells.
We have previously shown that microtubules in nonneuronal cells form long, stable bundles after transfection with the embryonic neuronal microtubule-associated protein MAP2c. In this study, we found that treating MAP2c-transfected cells with the actin depolymerising drug cytochalasin B led to the outgrowth of microtubule-containing processes from the cell surface. This effect was specific to MAP2c and did not occur in untransfected cells whose microtubules had been stabilised by treatment with taxol. The outgrowth and retraction of these processes during repeated cycles of cytochalasin addition and removal was followed by video time-lapse microscopy and was suggestive of a physical interaction between compressive forces exerted by the MAP2c-stabilised microtubule bundles and tensile forces originating in the cortical actin network. We suggest that MAP2c confers three properties on cellular microtubules that are essential for process outgrowth: stability, bundling and stiffness. The latter probably arises from the linking together of neighbouring tubulin subunits by three closely spaced tubulin-binding motifs in the MAP2 molecule that limits their motion relative to one another and thus reduces the flexibility of the polymer. Similar multimeric tubulin-binding domains in other proteins of the MAP2 class, including tau in axons and MAP4 in glial cells, may play the same role in the development and support of asymmetric cell morphology. Axial bundles of microtubules are found in growing neurites but not in growth cones, suggesting that the regulated expression of these MAP-induced properties makes an important contribution to the establishment of a stable process behind the advancing growth cone.
Friedrich Miescher Institute, Basel, Switzerland.
K
K
Edson
B
B
Weisshaar
A
A
Matus
D000199:Actins / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular; D003571:Cytochalasin B / Q000494:pharmacology*; D006801:Humans; D008113:Liver Neoplasms; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry*; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000648:ultrastructure; D014741:Video Recording
D000199:Actins; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; D011108:Polymers; D003571:Cytochalasin B