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Volume 271, Number 25, Issue of June 21, 1996 pp. 15153-15159
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Inhibition of Retinoic Acid Receptor Function and Retinoic Acid-regulated Gene Expression in Mouse Melanoma Cells by Calreticulin
A POTENTIAL PATHWAY FOR CYCLIC AMP REGULATION OF RETINOID ACTION

(Received for publication, February 22, 1996)

Dinakar Desai , Marek Michalak , Nishi K. Singh and Richard M. Niles

From the  Department of Biochemistry and Molecular Biology, Marshall University School of Medicine, Huntington, West Virginia 25755, the Medical Research Council Group in Molecular Biology of Membranes, Department of Biochemistry, University of Alberta, Edmonton, Canada T6G 2S2, and the Center for Biologics, Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

Calcium is a second messenger that controls a wide variety of cellular functions. Because of its multiple actions, there is a stringent requirement for calcium homeostasis, and this is achieved in part by a system of transport and storage proteins such as calreticulin located in the endoplasmic reticulum. Calreticulin is also found in the nucleus, suggesting that it may have a role in transcriptional regulation. It has been reported that calreticulin can inhibit steroid-regulated gene transcription by preventing receptor binding to DNA. Here we report that overexpression of the calreticulin gene in B16 mouse melanoma cells resulted in a decrease in retinoic acid (RA)-stimulated reporter gene expression. Gel shift analysis showed that purified calreticulin inhibited the binding of endogenous RAR to a -RA response element oligonucleotide, only if added prior to the addition of the oligonucleotide. Co-immunoprecipitation studies suggest a physical interaction between RAR and calreticulin. Transfection of the calreticulin gene into B16 cells inhibited the RA induction of protein kinase C, a marker of RA-induced differentiation. We also found that cyclic AMP increased the expression of calreticulin. Cyclic AMP may act to antagonize RA action by both decreasing RAR expression (Y. Xiao, D. Desai, T. Quick, and R. M. Niles, J. Cell Physiol., in press) and stimulating calreticulin levels.

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