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Phosphorylation of RGS14 by Protein Kinase A Potentiates Its Activity toward Gαi

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Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia 30322
Cite this: Biochemistry 2003, 42, 3, 811–819
Publication Date (Web):December 28, 2002
Copyright © 2003 American Chemical Society

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    Regulators of G protein signaling (RGS proteins) modulate Gα-directed signals because of the GTPase activating protein (GAP) activity of their conserved RGS domain. RGS14 and RGS12 are unique among RGS proteins in that they also regulate Gαi signals because of the guanine nucleotide dissociation inhibitor (GDI) activity of a GoLoco motif near their carboxy-termini. Little is known about cellular regulation of RGS proteins, although several are phosphorylated in response to G-protein directed signals. Here we show for the first time the phosphorylation of native and recombinant RGS14 in host cells. Direct stimulation of adenylyl cyclase or introduction of dibutyryl-cAMP induces phosphorylation of RGS14 in cells. This phosphorylation occurs through activation of cAMP-dependent protein kinase (PKA) since phosphate incorporation is completely blocked by a selective inhibitor of PKA but only partially or not at all blocked by inhibitors of other G-protein regulated kinases. We show that purified PKA phosphorylates two specific sites on recombinant RGS14, one of which, threonine 494 (Thr494), is immediately adjacent to the GoLoco motif. Because of this proximity, we focused on the possible effects of PKA phosphorylation on the GDI activity of RGS14. We found that mimicking phosphorylation on Thr494 enhanced the GDI activity of RGS14 toward Gαi nearly 3-fold, with no associated effect on the GAP activity toward either Gαi or Gαo. These findings implicate cAMP-induced phosphorylation as an important modulator of RGS14 function since phosphorylation could enhance RGS14 binding to Gαi−GDP, thereby limiting Gαi interactions with downstream effector(s) and/or enhancing Gβγ-dependent signals.

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     This research was supported by Grants NINDS R01NS37112 and NIGMS R01GM61849 awarded to J.R.H. from the National Institutes of Health.


     To whom correspondence should be addressed. Tel:  (404) 727-3641. Fax:  (404) 727-0365. Email:  [email protected].

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