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Volume 271, Number 41, Issue of October 11, 1996 pp. 25375-25381
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Inducible cAMP Early Repressor Can Modulate Tyrosine Hydroxylase Gene Expression after Stimulation of cAMP Synthesis

(Received for publication, April 30, 1996, and in revised form, July 25, 1996)

Cristina Tinti ^§ , Bruno Conti , Joseph F. Cubells , Kwang-Soo Kim , Harriet Baker and Tong H. Joh

From the  Laboratory of Molecular Neurobiology, Cornell University Medical College at The Burke Medical Research Institute, White Plains, New York 10605 and ^§ Dottorato in Medicina Sperimentale ed Applicata, Facoltá di Medicina e Chirurgia, Universitá degli Studi di Brescia, 25124 Brescia, Italy

Members of the CREB/CREM/ATF family of transcription factors either enhance or repress transcription after binding to the cAMP response elements (CREs) of numerous genes. The rat gene for tyrosine hydroxylase (TH) bears a canonical CRE, at base pairs 38 through 45 from the transcription initiation site, that is essential for basal and cAMP-stimulated transcription (Kim, K.-S., Lee, M. K., Carroll, J., and Joh, T. H. (1993) J. Biol. Chem. 268, 15689-15695; Lazaroff, M., Patankar, S., Yoon, S. O., and Chikaraishi, D. M. (1995) J. Biol. Chem. 270, 21579-21589). The current study identifies CRE-binding proteins induced in pharmacological paradigms characterized by TH activation.

PC12- and rat adrenal gland-derived nuclear proteins retarded a TH-CRE oligonucleotide in gel mobility shift assays with virtually identical patterns. These differed substantially from patterns exhibited by extracts from locus ceruleus or from neuroblastoma (SK-N-BE()C) and locus ceruleus-derived (CATH.a) cell lines. Forskolin stimulation of PC12 cells and reserpine treatment of rats increased, in nuclear extracts derived from cells and adrenal glands, respectively, the amount of a fast moving CRE/protein complex that was supershifted by an anti-CREM antibody. Subsequent Western, Northern, and polymerase chain reaction analyses indicated that a specific member of the CREM family, the inducible cAMP early repressor (ICER), was strongly induced in both systems. Cotransfection of PC12 cells with TH2400CAT plasmid and the expression vector pCMV-ICER-Ib demonstrated that ICER efficiently represses the transcriptional activity of the TH gene promoter. In addition, PKA-stimulated transcriptional activity of the promoter was effectively suppressed by ICER.

These results suggest that ICER can modulate cAMP-stimulated transcription of the TH gene and provide a model accounting for rapid reversal of increased TH transcription following elevations in cAMP.

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