Expression of chicken vinculin complements the adhesion-defective phenotype of a mutant mouse F9 embryonal carcinoma cell
| Autor: | R M Ezzell, D R Critchley, Jean-Luc Coll, Eileen D. Adamson, M Samuels, Timothy Cardozo |
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| Rok vydání: | 1993 |
| Předmět: |
Stress fiber
Embryonal Carcinoma Stem Cells Integrin Immunoblotting macromolecular substances Transfection Focal adhesion Mice Cell–cell interaction Cell Adhesion Animals Cloning Molecular Cell adhesion Actin Genetics biology Genetic Complementation Test Cell Biology Articles Vinculin Actins Cell biology Cytoskeletal Proteins Phenotype Mutation biology.protein Neoplastic Stem Cells Chickens |
| Zdroj: | The Journal of Cell Biology ResearcherID |
| ISSN: | 0021-9525 |
| Popis: | A mutant cell line, derived from the mouse embryonal carcinoma cell line F9, is defective in cell-cell adhesion (compaction) and in cell-substrate adhesion. We have previously shown that neither uvomorulin (E-cadherin) nor integrins are responsible for the mutant phenotype (Calogero, A., M. Samuels, T. Darland, S. A. Edwards, R. Kemler, and E. D. Adamson. 1991. Dev. Biol. 146:499-508). Several cytoskeleton proteins were assayed and only vinculin was found to be absent in mutant (5.51) cells. A chicken vinculin expression vector was transfected into the 5.51 cells together with a neomycin-resistance vector. Clones that were adherent to the substrate were selected in medium containing G418. Two clones, 5.51Vin3 and Vin4, were analyzed by Nomarski differential interference contrast and laser confocal microscopy as well as by biochemical and molecular biological techniques. Both clones adhered well to substrates and both exhibited F-actin stress fibers with vinculin localized at stress fiber tips in focal contacts. This was in marked contrast to 5.51 parental cells, which had no stress fibers and no vinculin. The mutant and complemented F9 cell lines will be useful models for examining the complex interactions between cytoskeletal and cell adhesion proteins. |
| Databáze: | OpenAIRE |
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