Hormones mobilize intracellular second messengers and initiate signalling cascades involving protein kinases and phosphatases, which are often spatially compartmentalized by anchoring proteins to increase signalling specificity1. These scaffold proteins may themselves be modulated by hormones2,3,4. In adipocytes, stimulation of β-adrenergic receptors increases cyclic AMP levels and activates protein kinase A (PKA)5, which stimulates lipolysis by phosphorylating hormone-sensitive lipase and perilipin6,7,8. Acute insulin treatment activates phosphodiesterase 3B, reduces cAMP levels and quenches β-adrenergic receptor signalling9. In contrast, chronic hyperinsulinaemic conditions (typical of type 2 diabetes) enhance β-adrenergic receptor-mediated cAMP production10. This amplification of cAMP signalling is paradoxical because it should enhance lipolysis, the opposite of the known short-term effect of hyperinsulinaemia. Here we show that in adipocytes, chronically high insulin levels inhibit β-adrenergic receptors (but not other cAMP-elevating stimuli) from activating PKA. We measured this using an improved fluorescent reporter and by phosphorylation of endogenous cAMP-response-element binding protein (CREB). Disruption of PKA scaffolding mimics the interference of insulin with β-adrenergic receptor signalling. Chronically high insulin levels may disrupt the close apposition of β-adrenergic receptors and PKA, identifying a new mechanism for crosstalk between heterologous signal transduction pathways.
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We thank Y. Ma for technical assistance and discussions and M. B. Yaffe for providing FHA cDNA. This work was supported by the Howard Hughes Medical Institute, the Alliance for Cell Signaling (R.Y.T.), the NIH (R.Y.T, C.J.H. and S.S.T) and the Johns Hopkins University School of Medicine and W. M. Keck Foundation (J.Z.). Author Contributions J.Z. and C.J.H. performed all the experiments; all authors contributed to planning the experiments, interpreting the data and writing the paper.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Supplementary Figures S1-S4, showing effect of insulin treatment on βAR-associated cAMP generation, effect of insulin treatment on PKA activity in the nucleus, effect of the functionally inactive Ht31p, and a model for insulin modulated compartmentation in adipocytes. (PDF 93 kb)
Additional information about the imaging methods used in this study. (DOC 24 kb)
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Zhang, J., Hupfeld, C., Taylor, S. et al. Insulin disrupts β-adrenergic signalling to protein kinase A in adipocytes. Nature 437, 569–573 (2005). https://doi.org/10.1038/nature04140
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