TY - JOUR
T1 - RGS12 and RGS14 GoLoco Motifs Are Gαi Interaction Sites with Guanine Nucleotide Dissociation Inhibitor Activity
AU - Kimple, Randall J.
AU - De Vries, Luc
AU - Tronchère, Hélène
AU - Behe, Cynthia I.
AU - Morris, Rebecca A.
AU - Farquhar, Marilyn Gist
AU - Siderovski, David P.
PY - 2001/8/3
Y1 - 2001/8/3
N2 - The regulators of G-protein signaling (RGS) proteins accelerate the intrinsic guanosine triphosphatase activity of heterotrimeric G-protein a subunits and are thus recognized as key modulators of G-protein-coupled receptor signaling. RGS12 and RGS14 contain not only the hallmark RGS box responsible for GTPase-accelerating activity but also a single Gα i/o-Loco (GoLoco) motif predicted to represent a second Gα interaction site. Here, we describe functional characterization of the GoLoco motif regions of RGS12 and RGS14. Both regions interact exclusively with Gαi1, Gαi2, and Gαi3 in their GDP-bound forms. In GTPγS binding assays, both regions exhibit guanine nucleotide dissociation inhibitor (GDI) activity, inhibiting the rate of exchange of GDP for GTP by Gαi1. Both regions also stabilize Gαi1 in its GDP-bound form, inhibiting the increase in intrinsic tryptophan fluorescence stimulated by AlF4-. Our results indicate that both RGS12 and RGS14 harbor two distinctly different Gα interaction sites: a previously recognized N-terminal RGS box possessing Gαi/o GAP activity and a C-terminal GoLoco region exhibiting Gαi GDI activity. The presence of two, independent Gα interaction sites suggests that RGS12 and RGS14 participate in a complex coordination of G-protein signaling beyond simple Gα GAP activity.
AB - The regulators of G-protein signaling (RGS) proteins accelerate the intrinsic guanosine triphosphatase activity of heterotrimeric G-protein a subunits and are thus recognized as key modulators of G-protein-coupled receptor signaling. RGS12 and RGS14 contain not only the hallmark RGS box responsible for GTPase-accelerating activity but also a single Gα i/o-Loco (GoLoco) motif predicted to represent a second Gα interaction site. Here, we describe functional characterization of the GoLoco motif regions of RGS12 and RGS14. Both regions interact exclusively with Gαi1, Gαi2, and Gαi3 in their GDP-bound forms. In GTPγS binding assays, both regions exhibit guanine nucleotide dissociation inhibitor (GDI) activity, inhibiting the rate of exchange of GDP for GTP by Gαi1. Both regions also stabilize Gαi1 in its GDP-bound form, inhibiting the increase in intrinsic tryptophan fluorescence stimulated by AlF4-. Our results indicate that both RGS12 and RGS14 harbor two distinctly different Gα interaction sites: a previously recognized N-terminal RGS box possessing Gαi/o GAP activity and a C-terminal GoLoco region exhibiting Gαi GDI activity. The presence of two, independent Gα interaction sites suggests that RGS12 and RGS14 participate in a complex coordination of G-protein signaling beyond simple Gα GAP activity.
UR - http://www.scopus.com/inward/record.url?scp=0035800830&partnerID=8YFLogxK
U2 - 10.1074/jbc.M103208200
DO - 10.1074/jbc.M103208200
M3 - Article
C2 - 11387333
AN - SCOPUS:0035800830
SN - 0021-9258
VL - 276
SP - 29275
EP - 29281
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 31
ER -