title: ADP-ribosylation factors modulate the cell surface transport of G protein-coupled receptors.
abstract: ADP-ribosylation factors (ARFs) regulate vesicular traffic through recruiting coat proteins. However, their functions in the anterograde transport of nascent G protein-coupled receptors (GPCRs) from the endoplasmic reticulum to the plasma membrane remain poorly explored. Here we show that treatment with brefeldin A, an inhibitor of guanine nucleotide exchange on ARFs, markedly attenuated the cell surface numbers of alpha(2B)-adrenergic receptor (AR), beta(2)-AR, angiotensin II type 1 receptor, and chemokine (CXC motif) receptor 4. Functional inhibition of individual ARF GTPases by transient expression of the GDP-bound, GTP-bound, and guanine nucleotide-deficient mutants showed that the five human ARFs differentially modulated receptor cell surface expression and that the ARF1 mutants produced the most profound inhibitory effect. Furthermore, expression of the ARF1 GTPase-activating protein (GAP) ARFGAP1 significantly blocked receptor transport. Interestingly, the GDP- and GTP-bound ARF1 mutants arrested the receptors in distinct intracellular compartments. Consistent with the reduced receptor cell surface expression, extracellular signal-regulated kinase 1 and 2 activation by receptor agonists was significantly attenuated by the GDP-bound mutant ARF1T31N. Moreover, coimmunoprecipitation showed that alpha(2B)-AR associated with ARF1 and glutathione transferase pull-down assay indicated that the alpha(2B)-AR C terminus directly interacted with ARF1. These data show that ARF1 GTPase is involved in the regulation of cell surface expression of GPCRs at multiple transport steps.
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Sciences Center, LA 70112, USA.
authors.forename: Chunmin
authors.initials: C
authors.lastname: Dong
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Zhang
authors.forename: Fuguo
authors.initials: F
authors.lastname: Zhou
authors.forename: Huijuan
authors.initials: H
authors.lastname: Dou
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Duvernay
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Guangyu
authors.initials: G
authors.lastname: Wu
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000502:physiology; D020727:ADP-Ribosylation Factors / Q000502:physiology*; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D021381:Protein Transport; D044140:Receptor, Angiotensin, Type 1 / Q000096:biosynthesis / Q000378:metabolism; D018341:Receptors, Adrenergic, alpha-2 / Q000096:biosynthesis / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000096:biosynthesis / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000096:biosynthesis / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: C547771:ADRA2B protein, human; C527276:CXCR4 protein, human; D044140:Receptor, Angiotensin, Type 1; D018341:Receptors, Adrenergic, alpha-2; D018343:Receptors, Adrenergic, beta-2; D019718:Receptors, CXCR4; D043562:Receptors, G-Protein-Coupled; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors