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Centre de Recherche en Cancérologie de l'Université Laval, L'Hôtel-Dieu de Québec, 11 Côte du Palais, Québec, G1R 2J6 Canada [J. H., F. H., J. L.], and Washington University School of Medicine, Section of Cancer Biology, St. Louis, Missouri [D. R. S.]
Phosphorylation of heat shock protein 27 (HSP27) has been suggested to play an important role in the regulation of F-actin dynamics in response to growth factors and stress. Because the microfilament network is one of the earliest targets of oxidative stress and because phosphorylation of HSP27 is strongly induced by reactive oxygen metabolites, we have investigated the role of HSP27 phosphorylation in regulating actin dynamics in response to oxidative stress. Experiments were done in Chinese hamster CCL39 cells overexpressing various levels of the wild-type or a nonphosphorylatable form of human HSP27 (pm3 HSP27). In control cells, both H2O2 and menadione induced fragmentation of F-actin, which forms aggregates and patches concentrated around the nucleus. Stable overexpression of wild-type HSP27, but not of pm3 HSP27, conferred resistance against actin fragmentation, suggesting that HSP27 has a phosphorylation-activated protective function against actin disruption by oxidative stress. Cell lines that overexpressed the highest levels of the wild-type form of human HSP27 also showed an increased cell survival following exposure to H2O2. In contrast, cells expressing pm3 HSP27 were as sensitive as the controls to the lethal effect of H2O2. These results suggest that phosphorylation of HSP27 is causally related to the regulation of microfilatment dynamics following oxidative stress and may be involved in mediating an adaptive response to oxyradical-generating agents such as carcinogens, anticancer drugs, and other xenobiotics.
1 This work was supported by La Société de Recherche sur le Cancer Inc (to J. L. and J. H.), Grant MT-7088 from the Medical Research Council of Canada (to J. L.), and NIH Grants HL 51469 and PO1-CA51116 (to D. R. S.).
2 To whom requests for reprints should be addressed.
Received 5/ 4/95. Accepted 11/ 6/95.
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