Metabolic Oxidative Stress-induced HSP70 Gene Expression Is Mediated through SAPK Pathway

doi:10.1074/jbc.273.45.29857

Metabolic Oxidative Stress-induced HSP70 Gene Expression Is Mediated through SAPK Pathway

ROLE OF Bcl-2 AND c-Jun NH₂-TERMINAL KINASE*

From the ^‡Department of Radiation Oncology, William Beaumont Hospital, Royal Oak, Michigan 48073 and the ^§Department of Molecular Biology and Research Center for Cell Differentiation, College of Natural Sciences, Seoul National University, Seoul 151-742, Korea

Abstract

In previous reports we demonstrated that glucose deprivation induces metabolic oxidative stress in drug-resistant human breast carcinoma MCF-7/ADR cells (Lee, Y. J., Galoforo, S. S., Berns, c. M., Chen, J. C., Davis, B. H., Swim, J. E., Corry, P. M., and Spitz, D. R. (1998)J. Biol. Chem. 273, 5294–5299). In the study described here, we investigated intracellular responses to metabolic oxidative stress. Northern blots show an increase in the level ofHSP70 and HSP28 mRNA in cells exposed to glucose-free medium for 1 h. One- and two-dimensional polyacrylamide gel analyses confirmed that glucose deprivation induced a family of HSPs, particularly an inducible HSP70. Overexpression of bcl-2 suppressed glucose deprivation-induced HSP70 gene expression, heat shock transcription factor-heat shock element binding activity, as well as c-Jun NH₂-terminal kinase (JNK1) activation. Expression of a dominant-negative mutant of JNK1 also suppressed glucose deprivation-induced JNK1 activation as well asHSP70 gene expression. Taken together, the stress-activated protein kinase signal transduction pathway is involved in glucose deprivation-induced heat shock gene expression.

Footnotes

↵* This work was supported by National Cancer Institute Grants CA48000 and CA44550, William Beaumont Hospital Research Institute Grant 97-06, the Korea Science and Engineering Foundation through the Research Center for Cell Differentiation, and the Seoul National University Research Fund.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¶ To whom all correspondence should be addressed: Dept. of Radiation Oncology, Research Laboratories, William Beaumont Hospital, 3601 W. Thirteen Mile Rd., Royal Oak, MI 48073. Tel.: 248-551-2568; Fax: 248-551-2443.
↵2 Y. J. Lee and P. M. Corry, unpublished data.
Abbreviations:

HSF

heat shock transcription factor

HSE

heat shock element

JNK

c-Jun NH₂-terminal kinase

SAPK

stress-activated protein kinase

PAGE

polyacrylamide gel electrophoresis

GSSG

oxidized glutathione

GSH

reduced glutathione

GAPDH

glyceraldehyde-3-phosphate dehydrogenase.
- Received June 29, 1998.
- Revision received August 20, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.