Abstract
Protein kinase A-dependent derepression of the human prodynorphin gene is regulated by the differential occupancy of the Dyn downstream regulatory element (DRE) site. Here, we show that a direct protein-protein interaction between DREAM and the CREM repressor isoform, alphaCREM, prevents binding of DREAM to the DRE and suggests a mechanism for cyclic AMP-dependent derepression of the prodynorphin gene in human neuroblastoma cells. Phosphorylation in the kinase-inducible domain of alphaCREM is not required for the interaction, but phospho-alphaCREM shows higher affinity for DREAM. The interaction with alphaCREM is independent of the Ca(2+)-binding properties of DREAM and is governed by leucine-charged residue-rich domains located in both alphaCREM and DREAM. Thus, our results propose a new mechanism for DREAM-mediated derepression that can operate independently of changes in nuclear Ca(2+).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs / genetics*
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Amino Acid Sequence
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Calcium / metabolism
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Calcium-Binding Proteins*
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Cell Line
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Colforsin / pharmacology
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Cyclic AMP Response Element Modulator
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Cyclic AMP Response Element-Binding Protein / metabolism
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Enkephalins / drug effects
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Enkephalins / genetics*
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Gene Expression Regulation / drug effects
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Genes, Regulator / genetics*
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Genes, Reporter / genetics
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Humans
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Kv Channel-Interacting Proteins
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Molecular Sequence Data
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Mutation / genetics
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Neuroblastoma
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Phosphorylation
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Protein Precursors / drug effects
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Protein Precursors / genetics*
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RNA, Messenger / metabolism
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Repressor Proteins / chemistry
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Repressor Proteins / genetics
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Repressor Proteins / metabolism*
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Sequence Alignment
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Transfection
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Tumor Cells, Cultured
Substances
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Calcium-Binding Proteins
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Cyclic AMP Response Element-Binding Protein
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DNA-Binding Proteins
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Enkephalins
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KCNIP3 protein, human
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Kv Channel-Interacting Proteins
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Protein Precursors
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RNA, Messenger
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Recombinant Proteins
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Repressor Proteins
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Cyclic AMP Response Element Modulator
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Colforsin
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preproenkephalin
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Cyclic AMP-Dependent Protein Kinases
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Calcium