title: β-Arrestin 1 inhibits the GTPase-activating protein function of ARHGAP21, promoting activation of RhoA following angiotensin II type 1A receptor stimulation.
abstract: Activation of the small GTPase RhoA following angiotensin II stimulation is known to result in actin reorganization and stress fiber formation. Full activation of RhoA, by angiotensin II, depends on the scaffolding protein β-arrestin 1, although the mechanism behind its involvement remains elusive. Here we uncover a novel partner and function for β-arrestin 1, namely, in binding to ARHGAP21 (also known as ARHGAP10), a known effector of RhoA activity, whose GTPase-activating protein (GAP) function it inhibits. Using yeast two-hybrid screening, a peptide array, in vitro binding studies, truncation analyses, and coimmunoprecipitation techniques, we show that β-arrestin 1 binds directly to ARHGAP21 in a region that transects the RhoA effector GAP domain. Moreover, we show that the level of a complex containing β-arrestin 1 and ARHGAP21 is dynamically increased following angiotensin stimulation and that the kinetics of this interaction modulates the temporal activation of RhoA. Using information gleaned from a peptide array, we developed a cell-permeant peptide that serves to inhibit the interaction of these proteins. Using this peptide, we demonstrate that disruption of the β-arrestin 1/ARHGAP21 complex results in a more active ARHGAP21, leading to less-efficient signaling via the angiotensin II type 1A receptor and, thereby, attenuation of stimulated stress fiber formation.
authors.affiliation: Neuroscience and Molecular Pharmacology, Division of Integrative Biology, IBLS, Wolfson Building, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: D F
authors.initials: DF
authors.lastname: Anthony
authors.forename: Y Y
authors.initials: YY
authors.lastname: Sin
authors.forename: S
authors.initials: S
authors.lastname: Vadrevu
authors.forename: N
authors.initials: N
authors.lastname: Advant
authors.forename: J P
authors.initials: JP
authors.lastname: Day
authors.forename: A M
authors.initials: AM
authors.lastname: Byrne
authors.forename: M J
authors.initials: MJ
authors.lastname: Lynch
authors.forename: G
authors.initials: G
authors.lastname: Milligan
authors.forename: M D
authors.initials: MD
authors.lastname: Houslay
authors.forename: G S
authors.initials: GS
authors.lastname: Baillie
mesh_terms: D000595:Amino Acid Sequence; D019390:Arrestins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D020690:GTPase-Activating Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D044140:Receptor, Angiotensin, Type 1 / Q000378:metabolism*; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism*; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: C459590:ARHGAP21 protein, human; C000605253:ARRB1 protein, human; D019390:Arrestins; D020690:GTPase-Activating Proteins; D019151:Peptide Library; D010455:Peptides; D044140:Receptor, Angiotensin, Type 1; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins; C066737:RHOA protein, human; D020742:rhoA GTP-Binding Protein