G protein-coupled receptors (GPCRs) constitute a super-family of cell surface receptors that regulate a variety of cell functions. Defective export trafficking of GPCRs from the endoplasmic reticulum (ER) through the Golgi to the cell surface is associated with the pathogenesis of a variety of human diseases, nephrogenic diabetes insipidus being one of the best-studied examples. However, the molecular mechanism underlying the export of GPCRs remains poorly understood. Our overall objective is to define the molecular mechanism of GPCR export trafficking and its functional role in regulating cellular responses to hormones and drugs. Under this broad objective, this proposal focuses on two important questions: 1) how GPCRs export from the ER? 2) How GPCRs transport from the ER to the Golgi? Our preliminary data have demonstrated that the motif consisting of a phenylalanine (F) and double leucine (L) spaced by six residues {F(x)6LL} is required for export of the alpha2B-adrenergic (AR) and angiotensin ll-type 1 (AT1R) receptors from the ER. This motif is highly conserved in many GPCRs and thus may provide a common mechanism for their export. Our data also indicate that different GPCRs may use different routes to move to the cell surface in neuroblastoma- glioma NG108 and human embryonic kidney HEK293 cell lines. Non-glycosylated alpha2B-AR uses a novel, as yet undefined pathway.
The Specific Aims are: 1) To define the mechanism of the F(x)6LL motif in mediating GPCR export from the ER. We will determine if the F(x)6LL motif is a common code for GPCR export from the ER, determine if the F(x)6LL motif functions as an ER export signal and/or regulates receptor folding, and define the structure-function relationship of the F(x)6LL motif. 2) To define the novel pathway for alpha2B-AR transport from the ER to the Golgi. We will define intracellular compartments and transport vesicles involved in alpha2B-AR transport and determine if glycosylation alters alpha2B-AR transport pathway. These studies will provide new and important information regarding the molecular mechanisms underlying GPCR export and its control on receptor function. Such information may help exploit the possibility of developing new therapeutic strategies for treating disease by targeting GPCR transport.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM076167-04
Application #
7615540
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Shapiro, Bert I
Project Start
2006-05-01
Project End
2010-04-30
Budget Start
2009-05-01
Budget End
2010-04-30
Support Year
4
Fiscal Year
2009
Total Cost
$241,983
Indirect Cost
Name
Louisiana State Univ Hsc New Orleans
Department
Pharmacology
Type
Schools of Medicine
DUNS #
782627814
City
New Orleans
State
LA
Country
United States
Zip Code
70112
Davis, Jason E; Xie, Xiayang; Guo, Jianhui et al. (2016) ARF1 promotes prostate tumorigenesis via targeting oncogenic MAPK signaling. Oncotarget 7:39834-39845
Zhang, Maoxiang; Davis, Jason E; Li, Chunman et al. (2016) GGA3 Interacts with a G Protein-Coupled Receptor and Modulates Its Cell Surface Export. Mol Cell Biol 36:1152-63
Zhou, Fuguo; Dong, Chunmin; Davis, Jason E et al. (2015) The mechanism and function of mitogen-activated protein kinase activation by ARF1. Cell Signal 27:2035-44
Lan, Tien-Hung; Wu, Guangyu; Lambert, Nevin A (2015) Lateral diffusion contributes to FRET from lanthanide-tagged membrane proteins. Biochem Biophys Res Commun 464:244-8
Wu, Guangyu; Davis, Jason E; Zhang, Maoxiang (2015) Regulation of α2B-Adrenerigc Receptor Export Trafficking by Specific Motifs. Prog Mol Biol Transl Sci 132:227-44
Zhu, Shu; Zhang, Maoxiang; Davis, Jason E et al. (2015) A single mutation in helix 8 enhances the angiotensin II type 1a receptor transport and signaling. Cell Signal 27:2371-9
Jang, Ji Hyun; Chun, Jung Nyeo; Godo, Shigeo et al. (2015) ROS and endothelial nitric oxide synthase (eNOS)-dependent trafficking of angiotensin II type 2 receptor begets neuronal NOS in cardiac myocytes. Basic Res Cardiol 110:21
Yang, Junjun; Yao, Wei; Qian, Guisheng et al. (2015) Rab5-mediated VE-cadherin internalization regulates the barrier function of the lung microvascular endothelium. Cell Mol Life Sci 72:4849-66
Lan, Tien-Hung; Liu, Qiuju; Li, Chunman et al. (2015) BRET evidence that β2 adrenergic receptors do not oligomerize in cells. Sci Rep 5:10166
Bertuccio, Claudia A; Lee, Shih-Liang; Wu, Guangyu et al. (2014) Anterograde trafficking of KCa3.1 in polarized epithelia is Rab1- and Rab8-dependent and recycling endosome-independent. PLoS One 9:e92013

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