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Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity
MDCK cells expressing RhoA or Rac1 mutants under control of the tetracycline repressible transactivator were used to examine short-term effects of known amounts of each mutant before, during, or after development of cell polarity. At low cell density, Rac1V12 cells had a flattened morphology and int...
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
1998
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2133034/ https://www.ncbi.nlm.nih.gov/pubmed/9660865 |
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author | Jou, Tzuu-Shuh James Nelson, W. |
author_facet | Jou, Tzuu-Shuh James Nelson, W. |
author_sort | Jou, Tzuu-Shuh |
collection | PubMed |
description | MDCK cells expressing RhoA or Rac1 mutants under control of the tetracycline repressible transactivator were used to examine short-term effects of known amounts of each mutant before, during, or after development of cell polarity. At low cell density, Rac1V12 cells had a flattened morphology and intact cell–cell contacts, whereas Rac1N17 cells were tightly compacted. Abnormal intracellular aggregates formed between Rac1N17, F-actin, and E-cadherin in these nonpolarized cells. At all subsequent stages of polarity development, Rac1N17 and Rac1V12 colocalized with E-cadherin and F-actin in an unusual beaded pattern at lateral membranes. In polarized cells, intracellular aggregates formed with Rac1V12, F-actin, and an apical membrane protein (GP135). At low cell density, RhoAV14 and RhoAN19 were localized in the cytoplasm, and cells were generally flattened and more fibroblastic than epithelial in morphology. In polarized RhoAV14 cells, F-actin was diffuse at lateral membranes and prominent in stress fibers on the basal membrane. GP135 was abnormally localized to the lateral membrane and in intracellular aggregates, but E-cadherin distribution appeared normal. In RhoAN19 cells, F-actin, E-cadherin, and GP135 distributions were similar to those in controls. Expression of either RhoAV14 or RhoAN19 in Rac1V12 cells disrupted Rac1V12 distribution and caused cells to adopt the more fibroblastic, RhoA mutant phenotype. We suggest that Rac1 and RhoA are involved in the transition of epithelial cells from a fibroblastic to a polarized structure and function by direct and indirect regulation of actin and actin-associated membrane protein organizations. |
format | Text |
id | pubmed-2133034 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1998 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21330342008-05-01 Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity Jou, Tzuu-Shuh James Nelson, W. J Cell Biol Articles MDCK cells expressing RhoA or Rac1 mutants under control of the tetracycline repressible transactivator were used to examine short-term effects of known amounts of each mutant before, during, or after development of cell polarity. At low cell density, Rac1V12 cells had a flattened morphology and intact cell–cell contacts, whereas Rac1N17 cells were tightly compacted. Abnormal intracellular aggregates formed between Rac1N17, F-actin, and E-cadherin in these nonpolarized cells. At all subsequent stages of polarity development, Rac1N17 and Rac1V12 colocalized with E-cadherin and F-actin in an unusual beaded pattern at lateral membranes. In polarized cells, intracellular aggregates formed with Rac1V12, F-actin, and an apical membrane protein (GP135). At low cell density, RhoAV14 and RhoAN19 were localized in the cytoplasm, and cells were generally flattened and more fibroblastic than epithelial in morphology. In polarized RhoAV14 cells, F-actin was diffuse at lateral membranes and prominent in stress fibers on the basal membrane. GP135 was abnormally localized to the lateral membrane and in intracellular aggregates, but E-cadherin distribution appeared normal. In RhoAN19 cells, F-actin, E-cadherin, and GP135 distributions were similar to those in controls. Expression of either RhoAV14 or RhoAN19 in Rac1V12 cells disrupted Rac1V12 distribution and caused cells to adopt the more fibroblastic, RhoA mutant phenotype. We suggest that Rac1 and RhoA are involved in the transition of epithelial cells from a fibroblastic to a polarized structure and function by direct and indirect regulation of actin and actin-associated membrane protein organizations. The Rockefeller University Press 1998-07-13 /pmc/articles/PMC2133034/ /pubmed/9660865 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Jou, Tzuu-Shuh James Nelson, W. Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title | Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title_full | Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title_fullStr | Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title_full_unstemmed | Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title_short | Effects of Regulated Expression of Mutant RhoA and Rac1 Small GTPases on the Development of Epithelial (MDCK) Cell Polarity |
title_sort | effects of regulated expression of mutant rhoa and rac1 small gtpases on the development of epithelial (mdck) cell polarity |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2133034/ https://www.ncbi.nlm.nih.gov/pubmed/9660865 |
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