title: Cell-type specific expression of a dominant negative PKA mutation in mice.
abstract: 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.
authors.affiliation: Department of Pharmacology, University of Washington, Seattle, Washington, United States of America.
authors.forename: Brandon S
authors.initials: BS
authors.lastname: Willis
authors.forename: Colleen M
authors.initials: CM
authors.lastname: Niswender
authors.forename: Thomas
authors.initials: T
authors.lastname: Su
authors.forename: Paul S
authors.initials: PS
authors.lastname: Amieux
authors.forename: G Stanley
authors.initials: GS
authors.lastname: McKnight
mesh_terms: D000483:Alleles; D000818:Animals; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000378:metabolism; D005799:Genes, Dominant; D005947:Glucose / Q000378:metabolism; D019426:Integrases / Q000235:genetics; D051379:Mice; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D017868:Cyclic AMP-Dependent Protein Kinases; C045073:Cre recombinase; D019426:Integrases; D005947:Glucose