Analysis of the proenkephalin second messenger-inducible enhancer in rat striatal cultures.

Konradi C, Cole RL, Green D, Senatus P, Leveque JC, Pollack AE, Grossbard SJ, Hyman SE
J Neurochem. 1995 65 (3): 1007-15

PMID: 7643080 · DOI:10.1046/j.1471-4159.1995.65031007.x

We have previously shown that in cell extracts from rat striatum, cyclic AMP response element (CRE) binding protein (CREB), rather than AP-1 proteins, preferentially interacts with the CRE-2 element of the proenkephalin second messenger-inducible enhancer, even under conditions in which AP-1 proteins are highly induced. Here we use primary striatal cultures to permit a more detailed analysis of CRE-2 function and protein binding in relevant neural cell types. By transfection we find that in primary striatal cultures, as in transformed cell lines, the CRE-1 and CRE-2 elements are required for significant induction by cyclic AMP. We report that cyclic AMP induction of the proenkephalin gene in striatal cultures is protein synthesis independent, excluding a role for newly synthesized proteins like c-Fos. We also show that cyclic AMP induces CREB phosphorylation and that phosphorylated CREB interacts strongly with CRE-2 and weakly with CRE-1. The predominant protein bound to CRE-1 is not CREB, however, and remains to be identified. Despite some prior predictions, we do not find a role for c-Fos in cyclic AMP regulation of proenkephalin gene expression in neurons.

MeSH Terms (22)

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone Adenosine Adenosine-5'-(N-ethylcarboxamide) Animals Base Sequence Cells, Cultured Colforsin Corpus Striatum Cyclic AMP Cyclic AMP Response Element-Binding Protein Enhancer Elements, Genetic Enkephalins Fetus Genes, fos Molecular Sequence Data Phosphorylation Protein Precursors Rats Rats, Sprague-Dawley RNA, Messenger Second Messenger Systems Transcription Factor AP-1

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