An in vivo assay for members of the cAMP response element-binding protein family of transcription factors.
The CREB/ATF family of dimeric basic/leucine zipper (bZIP) transcription factors have been implicated in signal transduction by several agents including intracellular cAMP. The ubiquity of the cAMP-signaling pathway in mammalian cells has made it difficult to assay individual family members in their native form and dissect their roles in signal transduction. In light of observations that ATF1 exhibits several cell-specific characteristics and is allosterically regulated, the above problem has represented a particular limitation for studying ATF1. To address this we have exploited the modular nature of the bZIP domain to design fusion proteins that closely resemble native ATF1 but have altered DNA binding and dimerization specificity. The activity of these novel proteins can be monitored following their introduction into a cAMP-responsive cell type. Thus we describe an assay for members of the CREB family that should be generally applicable in a range of cell types and to other bZIP protein families.
Ribeiro
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Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire, United Kingdom.
Brown
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Lee
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D000595:Amino Acid Sequence; D001483:Base Sequence; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D017362:Cyclic AMP Response Element-Binding Protein; D009838:Oligodeoxyribonucleotides; D000242:Cyclic AMP