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  • 1
    ISSN: 1432-2013
    Keywords: Key words Cell cycle ; Cytokinesis ; Heat shock protein 70 (HSP70) ; Temperature-sensitive mutant ; Thermotolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Cells of a temperature-sensitive mutant line (tsFT101) derived from a mouse mammary carcinoma cell line (FM3A) become multinucleated at a non-permissive temperature of 39°C because of disturbed cytokinesis. To explore how this relates to thermotolerance, we examined the proliferative activity of, and heat shock protein (HSP) expression in, FM3A and tsFT101 cells cultured at 37°C and 39°C after heat shock pretreatment (15 min exposure at 45°C). FM3A cells developed thermotolerance when cultured at both 37°C and 39°C, but whereas tsFT101 cells developed thermotolerance at 37°C, this was markedly reduced at 39°C. Western blot analysis showed similar degrees of expression of constitutive HSP70 (HSP73) in FM3A and tsFT101 cells after heat shock pretreatment at both 37°C and 39°C. However, expression of inducible HSP70 (HSP72) was reduced in tsFT101 cells at 39°C compared to 37°C and to FM3A cells at both 37°C and 39°C. Heat shock pretreatment activated DNA binding of heat shock transcription factor (HSF) in FM3A cells at 37°C and 39°C, but only at 37°C in tsFT101 cells. These results indicate that (1) multinucleation caused by disturbed cytokinesis increases temperature sensitivity, (2) HSP70 is critical for the development of thermotolerance in both FM3A and tsFT101 cells, and (3) decreased expression of inducible HSP70 parallels deficient development of thermotolerance in tsFT101 cells cultured at a non-permissive temperature.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Key words Cell division ; Cytokinesis ; G-actin ; F-actin ; Multinuclear cells ; Profilin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The temperature-sensitive mutant cells (tsFT101) derived from a mouse mammary carcinoma cell line, FM3A, become multinucleated at a non-permissive temperature of 39°C. To further understand the molecular mechanism of such cytokinetic disturbance, we examined the expression of profilin, the main regulator of the transition of globular actin (G-actin) to filamentous actin (F-actin). RT-PCR analysis of mouse profilin cDNA from tsFT101 showed a point mutation (177 A → G) which was a wobble mutation causing no change in the encoded amino acid. The expression level of profilin mRNA was, however, diminished in cultured tsFT101 cells under non-permissive temperatures compared with wild-type FM3A cells in association with multinucleation. A stable transfection of profilin cDNA expression vector to tsFT101 cells prevented multinuclear cell formation when cultured at 39°C. In contrast, antisense profilin cDNA expression vector did not alter multinuclear cell formation. The primary cause of the cytokinetic disturbance of tsFT101 cells may be due to the diminished level of profilin gene expression.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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