beta-Arrestin-mediated PDE4 cAMP phosphodiesterase recruitment regulates beta-adrenoceptor switching from Gs to Gi.

Baillie, George S

Sood, Arvind

McPhee, Ian

Gall, Irene

Perry, Stephen J

Lefkowitz, Robert J

Houslay, Miles D

United States

2013-09-05T15:37:01Z

2003-02-04

Phosphorylation of the beta(2) adrenoreceptor (beta(2)AR) by cAMP-activated protein kinase A (PKA) switches its predominant coupling from stimulatory guanine nucleotide regulatory protein (G(s)) to inhibitory guanine nucleotide regulatory protein (G(i)). beta-Arrestins recruit the cAMP-degrading PDE4 phosphodiesterases to the beta(2)AR, thus controlling PKA activity at the membrane. Here we investigate a role for PDE4 recruitment in regulating G protein switching by the beta(2)AR. In human embryonic kidney 293 cells overexpressing a recombinant beta(2)AR, stimulation with isoprenaline recruits beta-arrestins 1 and 2 as well as both PDE4D3 and PDE4D5 to the receptor and stimulates receptor phosphorylation by PKA. The PKA phosphorylation status of the beta(2)AR is enhanced markedly when cells are treated with the selective PDE4-inhibitor rolipram or when they are transfected with a catalytically inactive PDE4D mutant (PDE4D5-D556A) that competitively inhibits isoprenaline-stimulated recruitment of native PDE4 to the beta(2)AR. Rolipram and PDE4D5-D556A also enhance beta(2)AR-mediated activation of extracellular signal-regulated kinases ERK12. This is consistent with a switch in coupling of the receptor from G(s) to G(i), because the ERK12 activation is sensitive to both inhibitors of PKA (H89) and G(i) (pertussis toxin). In cardiac myocytes, the beta(2)AR also switches from G(s) to G(i) coupling. Treating primary cardiac myocytes with isoprenaline induces recruitment of PDE4D3 and PDE4D5 to membranes and activates ERK12. Rolipram robustly enhances this activation in a manner sensitive to both pertussis toxin and H89. Adenovirus-mediated expression of PDE4D5-D556A also potentiates ERK12 activation. Thus, receptor-stimulated beta-arrestin-mediated recruitment of PDE4 plays a central role in the regulation of G protein switching by the beta(2)AR in a physiological system, the cardiac myocyte.

https://www.ncbi.nlm.nih.gov/pubmed/12552097

262787199

0027-8424

https://hdl.handle.net/10161/7803

eng

Proceedings of the National Academy of Sciences

Proc Natl Acad Sci U S A

10.1073/pnas.262787199

3',5'-Cyclic-AMP Phosphodiesterases

Animals

Animals, Newborn

Arrestins

Cell Line

Cells, Cultured

Cyclic AMP

Cyclic AMP-Dependent Protein Kinases

Cyclic Nucleotide Phosphodiesterases, Type 3

Cyclic Nucleotide Phosphodiesterases, Type 4

Enzyme Activation

Enzyme Inhibitors

Genes, Dominant

Green Fluorescent Proteins

Humans

Isoquinolines

Luminescent Proteins

Models, Biological

Myocardium

Pertussis Toxin

Phosphoric Diester Hydrolases

Phosphorylation

Rats

Receptors, Adrenergic, beta

Rolipram

Signal Transduction

Sulfonamides

Time Factors

Transfection

beta-Arrestins

beta-Arrestin-mediated PDE4 cAMP phosphodiesterase recruitment regulates beta-adrenoceptor switching from Gs to Gi.

Journal article

https://www.ncbi.nlm.nih.gov/pubmed/12552097

940

945

3

Basic Science Departments

Biochemistry

Chemistry

Clinical Science Departments

Duke

Duke Cancer Institute

Institutes and Centers

Medicine

Medicine, Cardiology

Pathology

School of Medicine

Trinity College of Arts & Sciences

Published

100