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Identification of a Cyclic-AMP-Responsive Element within the Rat Somatostatin Gene
Marc R. Montminy, Kevin A. Sevarino, John A. Wagner, Gail Mandel and Richard H. Goodman
Proceedings of the National Academy of Sciences of the United States of America
Vol. 83, No. 18 (Sep. 15, 1986), pp. 6682-6686
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/28235
Page Count: 5
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We have examined the regulation of somatostatin gene expression by cAMP in PC12 rat pheochromocytoma cells transfected with the rat somatostatin gene. Forskolin at 10 μ M caused a 4-fold increase in somatostatin mRNA levels within 4 hr of treatment in stably transfected cells. Chimeric genes containing the somatostatin gene promoter fused to the bacterial reporter gene encoding chloramphenicol acetyltransferase were also induced by cAMP in PC12 cells. To delineate the sequences required for response to cAMP, we constructed a series of promoter deletion mutants. Our studies defined a region between 60 and 29 base pairs upstream from the transcriptional initiation site that conferred cAMP responsiveness when placed adjacent to the simian virus 40 promoter. Within the cAMP-responsive element of the somatostatin gene, we observed an 8-base palindrome, 5′-TGACGTCA-3′, which is highly conserved in many other genes whose expression is regulated by cAMP. cAMP responsiveness was greatly reduced when the somatostatin fusion genes were transfected into the mutant PC12 line A126-1B2, which is deficient in cAMP-dependent protein kinase 2. Our studies indicate that transcriptional regulation of the somatostatin gene by cAMP requires protein kinase 2 activity and may depend upon a highly conserved promoter element.
Proceedings of the National Academy of Sciences of the United States of America © 1986 National Academy of Sciences