title: Core engagement with β-arrestin is dispensable for agonist-induced vasopressin receptor endocytosis and ERK activation.
abstract: G protein-coupled receptors (GPCRs) exhibit highly conserved activation and signaling mechanisms by which agonist stimulation leads to coupling of heterotrimeric G proteins and generation of second messenger response. This is followed by receptor phosphorylation, primarily in the carboxyl terminus but also in the cytoplasmic loops, and subsequent binding of arrestins. GPCRs typically recruit arrestins through two different sets of interactions, one involving phosphorylated receptor tail and the other mediated by the receptor core. The engagement of both set of interactions (tail and core) is generally believed to be necessary for arrestin-dependent functional outcomes such as receptor desensitization, endocytosis, and G protein-independent signaling. Here we demonstrate that a vasopressin receptor (V2R) mutant with truncated third intracellular loop (V2RΔICL3) can interact with β-arrestin 1 (βarr1) only through the phosphorylated tail without engaging the core interaction. Of interest, such a partially engaged V2RΔICL3-βarr1 complex can efficiently interact with clathrin terminal domain and ERK2 MAPK in vitro. Furthermore, this core interaction-deficient V2R mutant exhibits efficient endocytosis and ERK activation upon agonist stimulation. Our data suggest that core interaction with βarr is dispensable for V2R endocytosis and ERK activation and therefore provide novel insights into refining the current understanding of functional requirements in biphasic GPCR-βarr interaction.
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India.
authors.forename: Punita
authors.initials: P
authors.lastname: Kumari
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India.
authors.forename: Eshan
authors.initials: E
authors.lastname: Ghosh
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India.
authors.forename: Ravi
authors.initials: R
authors.lastname: Ranjan
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, UP 226031, India.
authors.forename: Shalini
authors.initials: S
authors.lastname: Dogra
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, UP 226031, India.
authors.forename: Prem N
authors.initials: PN
authors.lastname: Yadav
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India arshukla@iitk.ac.in.
authors.forename: Arun K
authors.initials: AK
authors.lastname: Shukla
mesh_terms: D000595:Amino Acid Sequence; D019390:Arrestins / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020935:MAP Kinase Signaling System; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000378:metabolism*; D000071559:beta-Arrestin 1 / Q000378:metabolism*
chemical_list: C000605253:ARRB1 protein, human; D019390:Arrestins; D002966:Clathrin; D043562:Receptors, G-Protein-Coupled; D017483:Receptors, Vasopressin; D000071559:beta-Arrestin 1; D019950:Mitogen-Activated Protein Kinase 1; D019204:GTP-Binding Proteins