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J Biol Chem, Vol. 273, Issue 43, 27904-27910, October 23, 1998

Metallothionein Induction in Response to Restraint Stress
TRANSCRIPTIONAL CONTROL, ADAPTATION TO STRESS, AND ROLE OF GLUCOCORTICOID

Kalpana Ghoshal, Yijie Wang, John F. Sheridan^§¶, and Samson T. Jacob^§

From the Departments of  Medical Biochemistry and  Medical Microbiology and Immunology and the ^§ Comprehensive Cancer Center, Ohio State University College of Medicine and the ^¶ Section of Oral Biology, Ohio State University College of Dentistry, Columbus, Ohio 43210

Metallothioneins (MT) have been implicated in the protection of cells from oxidative stress. We studied the molecular mechanism of induction of MT-I and MT-II in response to restraint stress using a mouse model system in which the animals were restrained in well ventilated polypropylene tubes for 12 h each day (one cycle). Here, we show that MT-I and MT-II mRNA levels were elevated as much as 10-20-fold after just one cycle of this simple stress. Stress-mediated MT induction occurred at the transcriptional level. The level of MT mRNA correlated with the stress-induced increase, and not with the diurnal variation, in the level of serum glucocorticoid. Treatment of the mice with RU 486, a glucocorticoid receptor antagonist, prior to restraint stress inhibited MT induction by at least 50%. Furthermore, the glucocorticoid responsive element-binding activity in the liver nuclear extracts from the stressed mice was significantly higher than that in the control mice. The complex formations between the transcription factor Sp1, MTF1, or MLTF/ARE and the respective specific oligonucleotides were not altered in the liver from the stressed mouse. The MT mRNA levels returned to the basal level at the end of nine cycles of stress, indicating habituation of the animals to restraint stress. At this stage, exposure of the animals to another type of stress, treatment with heavy metals, resulted in further induction of MT. These data indicate that glucocorticoid is the primary physiological factor responsible for MT induction following restraint stress, and the glucocorticoid receptor is the major transcription factor involved in this process.

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