Expression of transfected mutant beta-actin genes: alterations of cell morphology and evidence for autoregulation in actin pools
| Autor: | U. Aebi, G Latter, John Leavitt, Madhu Varma, L Kedes, S Burbeck, Peter W. Gunning, Sun-Yu Ng |
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| Rok vydání: | 1987 |
| Předmět: |
Cell division
Mutant macromolecular substances Biology Transfection Cell morphology Microfilament Cell Line Homeostasis Humans Beta-actin Neoplastic transformation Cytoskeleton Molecular Biology Alleles Actin Cell Biology Molecular biology Actins Phenotype Gene Expression Regulation Genes Mutation Cell Division Research Article |
| Zdroj: | Molecular and Cellular Biology. 7:2457-2466 |
| ISSN: | 1098-5549 0270-7306 |
| Popis: | Two different mutant human beta-actin genes have been introduced into normal diploid human (KD) fibroblasts and their immortalized derivative cell line, HuT-12, to assess the impact of an abnormal cytoskeletal protein on cellular phenotypes such as morphology, growth characteristics, and properties relating to the neoplastic phenotype. A mutant beta-actin containing a single mutation (Gly-244----Asp-244) was stable and was incorporated into cytoskeletal stress fibers. Transfected KD cells which expressed the stable mutant beta-actin in excess of normal beta-actin were morphologically altered. In contrast, a second mutant beta-actin gene containing two additional mutations (Gly-36----Glu-36 and Glu-83----Asp-83, as well as Gly-244----Asp-244) did not alter cell morphology when expressed at high levels in transfected cells, but the protein was labile and did not accumulate in stress fibers. In both KD and HuT-12 cells, endogenous beta- and gamma-actin decreased in response to high-level expression of the stable mutant beta-actin, in a manner consistent with autoregulatory feedback of actin concentrations. Since the percent decreases in the endogenous beta- and gamma-actins were equal, the ratio of net beta-actin (mutant plus normal) to gamma-actin was significantly increased in the transfected cells. Antisera capable of distinguishing the mutant from the normal epitope revealed that the mutant beta-actin accumulated in stress fibers but did not participate in the formation of the actin filament-rich perinuclear network. These observations suggest that different intracellular locations differentially incorporate actin into cytoskeletal microfilaments. The dramatic impact on cell morphology and on beta-actin/gamma-actin ratios in the transfected diploid KD cells may be related to the acquisition of some of the characteristics of cells that underwent the neoplastic transformation event that originally led to the appearance of the beta-actin mutations. |
| Databáze: | OpenAIRE |
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