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Cell-Type Specific Expression of a Dominant Negative PKA Mutation in Mice

By Brandon S. Willis, Colleen M. Niswender, Thomas Su, Paul S. Amieux and G. Stanley McKnight


We employed the Cre recombinase/loxP system to create a mouse line in which PKA activity can be inhibited in any cell-type that expresses Cre recombinase. The mouse line carries a mutant Prkar1a allele encoding a glycine to aspartate substitution at position 324 in the carboxy-terminal cAMP-binding domain (site B). This mutation produces a dominant negative RIα regulatory subunit (RIαB) and leads to inhibition of PKA activity. Insertion of a loxP-flanked neomycin cassette in the intron preceding the site B mutation prevents expression of the mutant RIαB allele until Cre-mediated excision of the cassette occurs. Embryonic stem cells expressing RIαB demonstrated a reduction in PKA activity and inhibition of cAMP-responsive gene expression. Mice expressing RIαB in hepatocytes exhibited reduced PKA activity, normal fasting induced gene expression, and enhanced glucose disposal. Activation of the RIαB allele in vivo provides a novel system for the analysis of PKA function in physiology

Topics: Research Article
Publisher: Public Library of Science
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Provided by: PubMed Central

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  2. (1986). Characterization of the phosphoenolpyruvate carboxykinase (GTP) promoter-regulatory region. II. Identification of cAMP and glucocorticoid regulatory domains.
  3. (2004). Comparative PRKAR1A genotype-phenotype analyses in humans with Carney complex and prkar1a haploinsufficient mice.
  4. (1975). Comparison of adenosine 39:59-monophosphate-dependent protein kinases from rabbit skeletal and bovine heart muscle.
  5. (1997). Compensatory regulation of RIalpha protein levels in protein kinase A mutant mice.
  6. (2001). Control of hepatic gluconeogenesis through the transcriptional coactivator PGC-1.
  7. (2005). Cre recombinase-dependent expression of a constitutively active mutant allele of the catalytic subunit of protein kinase A.
  8. (2001). CREB regulates hepatic gluconeogenesis through the coactivator PGC-1.
  9. (1991). Cyclic AMP induction of phosphoenolpyruvate carboxykinase (GTP) gene transcription is mediated by multiple promoter elements.
  10. (1991). Cyclic AMP second messenger systems.
  11. (1989). Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133.
  12. (1992). Cyclic AMP-dependent protein kinase controls basal gene activity and steroidogenesis in Y1 adrenal tumor cells.
  13. (2003). Cytochrome-P450 phosphorylation as a functional switch.
  14. (1998). Defective motor behavior and neural gene expression in RIIbeta-protein kinase A mutant mice.
  15. (1995). Derivation of novel embryonic stem cell lines and targeting of cyclic AMP-dependent protein kinase genes. Recent Prog Horm
  16. (2000). DNA excision in liver by an albumin-Cre transgene occurs progressively with age.
  17. (1990). Dynamics of the distribution of cyclic AMP-dependent protein kinase in living cells.
  18. (1989). Expression and characterization of mutant forms of the type I regulatory subunit of cAMPdependent protein kinase. The effect of defective cAMP binding on holoenzyme activation.
  19. (2000). Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development.
  20. (1989). Functional characterization of cAMP-binding mutations in type I protein kinase.
  21. (2000). Genetic heterogeneity and spectrum of mutations of the PRKAR1A gene in patients with the carney complex.
  22. (1996). Genetically lean mice result from targeted disruption of the RII beta subunit of protein kinase A.
  23. (2006). Haploinsufficiency at the protein kinase A RI alpha gene locus leads to fertility defects in male mice and men.
  24. (1985). Hotspots for spontaneous and mutageninduced lesions in regulatory subunit of cyclic AMP-dependent protein kinase in S49 mouse lymphoma cells.
  25. (1986). Identification of a cyclic-AMP-responsive element within the rat somatostatin gene.
  26. (2007). Impaired regulation of hepatic glucose production in mice lacking the forkhead transcription factor Foxo1 in liver.
  27. (2002). Increased basal cAMP-dependent protein kinase activity inhibits the formation of mesoderm-derived structures in the developing mouse embryo.
  28. (1987). Inhibition of intracellular cAMP-dependent protein kinase using mutant genes of the regulatory type I subunit.
  29. (2006). Inhibition of protein kinase A in murine enteric neurons causes lethal intestinal pseudoobstruction.
  30. (2007). Insulin modulates gluconeogenesis by inhibition of the coactivator TORC2.
  31. (2003). Insulinregulated hepatic gluconeogenesis through FOXO1-PGC-1alpha interaction.
  32. (2002). Liver repopulation by Bcl-x(L) transgenic hepatocytes.
  33. (2002). Molecular and behavioral effects of a null mutation in all PKA C beta isoforms.
  34. (1976). Molecular mechanisms of cyclic AMP action: a genetic approach. Recent Prog Horm
  35. (1992). Molecular physiology of the regulation of hepatic gluconeogenesis and glycolysis.
  36. (1994). Multiple pathway signal transduction by the cAMP-dependent protein kinase.
  37. (2002). Mutation of the Calpha subunit of PKA leads to growth retardation and sperm dysfunction.
  38. (2000). Mutations in the protein kinase A R1alpha regulatory subunit cause familial cardiac myxomas and Carney complex.
  39. (2000). Mutations of the gene encoding the protein kinase A type I-alpha regulatory subunit in patients with the Carney complex.
  40. (2003). Peroxisome proliferator-activated receptorgamma coactivator 1 alpha (PGC-1 alpha): transcriptional coactivator and metabolic regulator.
  41. (2000). Phosphoenolpyruvate carboxykinase is necessary for the integration of hepatic energy metabolism.
  42. (1988). Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB.
  43. (1989). Regulation of transcription by cyclic AMP-dependent protein kinase.
  44. (2009). Specific recruitment of protein kinase A to the immunoglobulin locus regulates classswitch recombination.
  45. (1978). Studies on the properties and mode of action of the purified regulatory subunit of bovine heart adenosine 39:59-monophosphate-dependent protein kinase.
  46. (2005). The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism.
  47. (1989). The expression of cAMP-dependent protein kinase subunits is differentially regulated during liver regeneration.
  48. (1997). The role of HNF1alpha, HNF3gamma, and cyclic AMP in glucose-6-phosphatase gene activation.
  49. (1989). Transcriptional induction of glucokinase gene by insulin in cultured liver cells A Dominant Negative PKA Mutation in
  50. (1988). Transcriptional regulation of a plasminogen activator gene by cyclic AMP in a homologous cell-free system. Involvement of cyclic AMP-dependent protein kinase in transcriptional control.
  51. (1997). Two novel brain-specific splice variants of the murine Cbeta gene of cAMP-dependent protein kinase.
  52. (1997). Type II regulatory subunits are not required for the anchoring-dependent modulation of Ca2+ channel activity by cAMP-dependent protein kinase.