title: Galpha16 activates Ras by forming a complex with tetratricopeptide  repeat 1 (TPR1) and  Son of Sevenless (SOS).
abstract: Many G protein-coupled receptors (GPCRs) are known to modulate cell growth and differentiation by stimulating the extracellular signal-regulated protein kinases (ERKs). In growth factor signaling, ERKs are typically stimulated through an elaborate network of modules consisting of adaptors, protein kinases, and the small GTPase Ras. The mechanism by which G protein signals tap into the ERK signaling pathway has thus far remain elusive. Members of the Gq family of G proteins, in particular Galpha16, have been shown to associate with tetratricopeptide repeat 1 (TPR1), an adaptor protein which preferentially binds to Ras. Here, we examined if TPR1 is indeed the missing link between Galpha16 signaling and Ras activation. Expression of Galpha16QL, a constitutively active mutant of Galpha16, in HEK 293 cells led to the formation of GTP-bound Ras and the subsequent phosphorylation of ERK. Likewise, stimulation of endogenou G16-coupled CCR1 chemokine receptors produced the same responses in human erythroleukemia cells. siRNA-mediated knockdown of TPR1 or expression of a dominant negative mutant of TPR1 effectively abolished the ability of Galpha16QL to induce Ras activation in HEK 293 cells. In contrast, these manipulations had no inhibitory effect on Galpha16QL induced activation of phospholipase Cbeta. Galpha16QL-induced phosphorylations of downstream targets including ERK, signal transducer and activator of transcription 3, and IkappaB kinase were significantly suppressed upon expression of the dominant negative mutant of TPR1. Furthermore, SOS2, a Ras guanine nucleotide exchange factor, was found to form a complex with TPR1 and Galpha16QL. Expression of SOS2 enhanced Galpha16QL-induced Ras activation and its subsequent signaling. Collectively, our results suggest that Galpha16 regulates multiple signaling pathways by activating Ras through its association with TPR1, but TPR1 is not required for Galpha16 to stimulate phospholipase Cbeta.
authors.affiliation: Department of Biochemistry, Molecular Neuroscience Center and Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
authors.forename: Andrew M F
authors.initials: AM
authors.lastname: Liu
authors.forename: Rico K H
authors.initials: RK
authors.lastname: Lo
authors.forename: Maggie M K
authors.initials: MM
authors.lastname: Lee
authors.forename: Yingchun
authors.initials: Y
authors.lastname: Wang
authors.forename: Wendy W S
authors.initials: WW
authors.lastname: Yeung
authors.forename: Maurice K C
authors.initials: MK
authors.lastname: Ho
authors.forename: Yan
authors.initials: Y
authors.lastname: Su
authors.forename: Richard D
authors.initials: RD
authors.lastname: Ye
authors.forename: Yung H
authors.initials: YH
authors.lastname: Wong
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D002460:Cell Line; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000235:genetics / Q000378:metabolism*; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D054799:Phospholipase C beta / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism*; D015398:Signal Transduction; D020664:Son of Sevenless Proteins / Q000378:metabolism*; D020703:ras GTPase-Activating Proteins / Q000378:metabolism
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C485509:SOS2 protein, human; D020664:Son of Sevenless Proteins; C475144:TTC1 protein, human; D020703:ras GTPase-Activating Proteins; D054799:Phospholipase C beta; C112631:G protein alpha 16; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins; D016283:Proto-Oncogene Proteins p21(ras)