Proteomic identification of Hsc70 as a mediator of RGS9-2 degradation by in vivo interactome analysis.
Changes in interactions between signaling proteins underlie many cellular functions. In the mammalian nervous system, a member of the Regulator of G protein Signaling family, RGS9-2 (Regulator of G protein Signaling, type 9), is a key regulator of dopamine and opioid signaling pathways that mediate motor control and reward behavior. Dynamic association of RGS9-2 with a neuronal protein R7BP (R7 family Binding Protein) has been found to be critically important for the regulation of the expression level of the complex by proteolytic mechanisms. Changes in RGS9-2 expression are observed in response to a number of signaling events and are thought to contribute to the plasticity of the neurotransmitter action. In this study, we report an identification of molecular chaperone Hsc70 (Heat shock cognate protein 70) as a critical mediator of RGS9-2 expression that is specifically recruited to the intrinsically disordered C-terminal domain of RGS9-2 following its dissociation from R7BP. Hsc70 was identified by a novel application of the quantitative proteomics approach developed to monitor interactome dynamics in mice using a set of controls contributed by knockout strains. We propose this application to be a useful tool for studying the dynamics of protein assemblies in complex models, such as signaling in the mammalian nervous system.
Posokhova
Ekaterina
E
Department of Pharmacology, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Uversky
Vladimir
V
Martemyanov
Kirill A
KA
D000818:Animals; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D002460:Cell Line; D054022:Cerebrum / Q000378:metabolism; D050883:HSC70 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007553:Isotope Labeling; D051379:Mice; D018345:Mice, Knockout; D025941:Protein Interaction Mapping / Q000379:methods; D040901:Proteomics / Q000379:methods*; D020710:RGS Proteins / Q000737:chemistry / Q000378:metabolism*
D050883:HSC70 Heat-Shock Proteins; C499440:R7BP protein, mouse; D020710:RGS Proteins; C116925:regulator of g-protein signaling 9