Abstract
Targeted disruption of the RIIbeta subunit of protein kinase A (PKA) produces lean mice that resist diet-induced obesity. In this report we examine the effects of the RIIbeta knockout on white adipose tissue physiology. Loss of RIIbeta is compensated by an increase in the RIalpha isoform, generating an isoform switch from a type II to a type I PKA. Type I holoenzyme binds cAMP more avidly and is more easily activated than the type II enzyme. These alterations are associated with increases in both basal kinase activity and the basal rate of lipolysis, possibly contributing to the lean phenotype. However, the ability of both beta(3)-selective and nonspecific beta-adrenergic agonists to stimulate lipolysis is markedly compromised in mutant white adipose tissue. This defect was found in vitro and in vivo and does not result from reduced expression of beta-adrenergic receptor or hormone-sensitive lipase genes. In contrast, beta-adrenergic stimulated gene transcription remains intact, and the expression of key genes involved in lipid metabolism is normal under both fasted and fed conditions. We suggest that the R subunit isoform switch disrupts the subcellular localization of PKA that is required for efficient transduction of signals that modulate lipolysis but not for those that mediate gene expression.
Highlights
Protein kinase A (PKA)1 transduces the cAMP-mediated signals from more than 30 different hormones and neurotransmitters, many of which may act simultaneously on a given cell to provoke discrete biological responses [1]
Because PKA signaling anomalies in RII mutant mice could theoretically arise from changes in the number of R or C subunits, rather than from the isoform switch, we quantified these proteins in white adipose tissue (WAT)
Whereas RI␣ compensation is virtually complete in mutant brown adipose tissue (BAT) [4], there is a 30% loss of R subunits overall in mutant WAT, as assessed by total cAMP-binding capacity
Summary
Protein kinase A (PKA)1 transduces the cAMP-mediated signals from more than 30 different hormones and neurotransmitters, many of which may act simultaneously on a given cell to provoke discrete biological responses [1]. In order to determine the cAMP affinities of PKA from wild type versus RII mutant WAT, the cAMP-binding capacity of tissue homogenates was measured.
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