title: G-protein signaling modulator-3 regulates heterotrimeric G-protein dynamics through dual association with Gβ and Gαi protein subunits.
abstract: Regulation of the assembly and function of G-protein heterotrimers (Gα·GDP/Gβγ) is a complex process involving the participation of many accessory proteins. One of these regulators, GPSM3, is a member of a family of proteins containing one or more copies of a small regulatory motif known as the GoLoco (or GPR) motif. Although GPSM3 is known to bind Gα(i)·GDP subunits via its GoLoco motifs, here we report that GPSM3 also interacts with the Gβ subunits Gβ1 to Gβ4, independent of Gγ or Gα·GDP subunit interactions. Bimolecular fluorescence complementation studies suggest that the Gβ-GPSM3 complex is formed at, and transits through, the Golgi apparatus and also exists as a soluble complex in the cytoplasm. GPSM3 and Gβ co-localize endogenously in THP-1 cells at the plasma membrane and in a juxtanuclear compartment. We provide evidence that GPSM3 increases Gβ stability until formation of the Gβγ dimer, including association of the Gβ-GPSM3 complex with phosducin-like protein PhLP and T-complex protein 1 subunit eta (CCT7), two known chaperones of neosynthesized Gβ subunits. The Gβ interaction site within GPSM3 was mapped to a leucine-rich region proximal to the N-terminal side of its first GoLoco motif. Both Gβ and Gα(i)·GDP binding events are required for GPSM3 activity in inhibiting phospholipase-Cβ activation. GPSM3 is also shown in THP-1 cells to be important for Akt activation, a known Gβγ-dependent pathway. Discovery of a Gβ/GPSM3 interaction, independent of Gα·GDP and Gγ involvement, adds to the combinatorial complexity of the role of GPSM3 in heterotrimeric G-protein regulation.
authors.affiliation: Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7365, USA.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Giguère
authors.forename: Geneviève
authors.initials: G
authors.lastname: Laroche
authors.forename: Emily A
authors.initials: EA
authors.lastname: Oestreich
authors.forename: David P
authors.initials: DP
authors.lastname: Siderovski
mesh_terms: D000818:Animals; D019556:COS Cells; D056404:Chaperonin Containing TCP-1 / Q000235:genetics / Q000378:metabolism; D002522:Chlorocebus aethiops; D004789:Enzyme Activation / Q000502:physiology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000235:genetics / Q000378:metabolism*; D044387:GTP-Binding Protein beta Subunits / Q000235:genetics / Q000378:metabolism*; D020730:Guanine Nucleotide Dissociation Inhibitors / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D054799:Phospholipase C beta / Q000235:genetics / Q000378:metabolism
chemical_list: C533333:CCT7 protein, human; C487218:GPSM3 protein, human; D044387:GTP-Binding Protein beta Subunits; D020730:Guanine Nucleotide Dissociation Inhibitors; D046912:Multiprotein Complexes; D054799:Phospholipase C beta; D056404:Chaperonin Containing TCP-1; D019206:GTP-Binding Protein alpha Subunits, Gi-Go