title: [Regulation of G protein-coupled receptor function by its binding proteins].
abstract: G protein-coupled receptors (GPCRs) are seven transmembrane receptors with an N-terminus in the extracellular region and C-terminus in the intracellular region. When an agonist binds to a GPCR, a signal is transduced into a cell through the activation of trimeric G proteins. Recently, it has been shown that the activities of GPCRs are regulated by multiple mechanisms. One of the mechanisms is regulation through the binding proteins to the carboxy (C)-terminus of GPCRs. In the present study, the binding partners for the C-terminus of the parathyroid hormone receptor (PTHR) and thromboxane A(2) receptor (TP) were searched for using yeast two-hybrid screening, and the functions of these proteins were investigated. We identified t-complex testis expressed-1 (Tctex-1) and 4.1G as associated proteins for the PTHR. Tctex-1 is one of the light chains of cytoplasmic dynein, which is a motor protein across microtubles. We found that Tctex-1 was involved in agonist-induced internalization of the PTHR. 4.1G, a cytoskeletal protein, facilitated the cell surface localization of the PTHR and augmented PHTR-mediated signal transduction. TPs consists of two splicing variants, TPalpha and TPbeta. As a result of yeast two-hybrid screening, two proteasomal proteins, proteasome activator PA28gamma and proteasome subunit alpha7, were identified as direct interacting proteins for TPbeta. TPbeta has a tendency to be retained in the intracellular compartment, probably due to its binding to proteasomes. We also demonstrated that TPalpha and TPbeta formed heterodimers, and the signal transduction through TPalpha was reduced by the formation of heterodimers. In conclusion, the proteins bound to GPCRs may regulate the intracellular traffic of GPCRs.
authors.affiliation: Department of Cellular Signaling, Graduate School of Pharmaceutical Sciences, Tohoku University, Aramaki, Aoba-ku, Sendai City, Japan. nakahata@mail.pharma.tohoku.ac.jp
authors.forename: Norimichi
authors.initials: N
authors.lastname: Nakahata
authors.forename: Masaki
authors.initials: M
authors.lastname: Saito
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000502:physiology; D004398:Dyneins; D019204:GTP-Binding Proteins / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000502:physiology; D009687:Nuclear Proteins / Q000502:physiology; D046988:Proteasome Endopeptidase Complex / Q000502:physiology; D011485:Protein Binding; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000737:chemistry / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000502:physiology*; D044244:Receptors, Thromboxane A2, Prostaglandin H2 / Q000502:physiology*; D015398:Signal Transduction; D056427:t-Complex Genome Region
chemical_list: D001324:Autoantigens; D003598:Cytoskeletal Proteins; C533388:Dynlt1 protein, rat; C480491:Epb41l2 protein, rat; C099558:Ki antigen; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D044168:Receptor, Parathyroid Hormone, Type 1; D043562:Receptors, G-Protein-Coupled; D044244:Receptors, Thromboxane A2, Prostaglandin H2; C488662:PSMA7 protein, human; D046988:Proteasome Endopeptidase Complex; D019204:GTP-Binding Proteins; D004398:Dyneins