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Ablation of stimulation of a cAMP-responsive promoter in CHO cell lines defective in their cAMP-dependent protein kinase system

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Somatic Cell and Molecular Genetics

Abstract

We have studied the requirement for an intact cAMP-dependent protein kinase (PKA) system to regulate cAMP-mediated gene transcription in Chinese hamster ovary (CHO) cells. Wild-type CHO cells and mutant CHO cell lines selected for their resistance to the growth inhibitory effect of 8-Br-cAMP and defective in their PKA system were transiently transfected with reporter plasmids containing 2.5 and 3.0 kb of the 5′-flanking sequence of the rat tyrosine aminotransferase (TAT) gene promoter. This segment of DNA contains no CRE-like sequences, yet wild-type transfectants exhibited a specific increase in TAT promoter activity following growth in medium containing 8-Br-cAMP. In CHO cell lines defective in their PKA, the transfected TAT promoter failed to respond to cAMP treatment. We conclude that an intact PKA system is necessary for the cAMP-mediated increase in TAT promoter activity in CHO cells and that there is no requirement for a CRE to see this effect.

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Authors and Affiliations

  1. Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, 20892, Bethesda, Maryland

    Robert D. Fleischmann, Clifford Jeng & Michael M. Gottesman

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  1. Robert D. Fleischmann
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  2. Clifford Jeng
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  3. Michael M. Gottesman
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Fleischmann, R.D., Jeng, C. & Gottesman, M.M. Ablation of stimulation of a cAMP-responsive promoter in CHO cell lines defective in their cAMP-dependent protein kinase system. Somat Cell Mol Genet 18, 103–111 (1992). https://doi.org/10.1007/BF01233157

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  • DOI: https://doi.org/10.1007/BF01233157

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