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J. Biochem, 1995, Vol. 118, No. 3 629-634
© 1995 Japanese Biochemical Society

other

Modulation of the Stress-Induced Synthesis of Stress Proteins by a Phorbol Ester and Okadaic Acid1

Hidenori Ito, Kaori Hasegawa, Yutaka Inaguma, Osamu Kozawa, Tomiko Asano and Kanefusa Kato2

Department of Biochemistry, Institute for Developmental Research, Aichi Human Service Center Kasugai, Aichi 480-03

2To whom correspondence should be addressed.

The expression of {alpha}B crystallin, hsp27, and hsp70 in C6 cells increased when the cells were exposed to arsenite (50{diaeresis} for 1h) or heat (42°C for 30min), as detected by specific immunoassays, Western blot analysis, and Northern blot analysis. When cells were exposed to arsenite in the presence of 0.1 {diaeresis}M phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, or 0.2 {diaeresis}okadaic acid, an inhibitor of phosphoserine/phosphothreonine protein phosphatases, expression of {alpha}B crystallin was markedly enhanced. The induction of hsp27 and hsp70 expression was also stimulated to a considerable extent in the same cells. The stimulatory effect of PMA was further enhanced in the presence of okadaic acid, but it was strongly inhibited in the presence of 0.5 {alpha}M staurosporine, an inhibitor of protein kinase C. PMA and okadaic acid also stimulated the response to heat stress of the expression of {alpha}B crystallin, but they barely stimulated the response to heat stress of hsp27. The extent of stimulation of the arsenite-induced responses by PMA and okadaic acid was greater when the concentration of arsenite (Le. the magnitude of the stress) was relatively low (25-50 {diaeresis}M). The arsenite-induced release of arachidonic acid from cells was also stimulated in the presence of PMA and/or okadaic acid, and the stimlatory effects of PMA and okadaic acid on the arsenite-induced accumlation of {alpha}B crystallin and hsp27 were strongly suppressed by quinacrine, an inhibitor of phospholipase A2. These results suggest that the stimulatory effects of PMA and okadaic acid on the stress responses are caused, in part, by the increased metabolic activity of the arachidonic acid cascade, as a consequence of the activation of phospholipase A2.

1This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science and Culture of Japan.


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 J. Biol. Chem.Home page
K. Kato, H. Ito, Y. Inaguma, K. Okamoto, and S. Saga
Synthesis and Accumulation of alpha B Crystallin in C6 Glioma Cells Is Induced by Agents That Promote the Disassembly of Microtubules
J. Biol. Chem., October 25, 1996; 271(43): 26989 - 26994.
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