title: Structure of the Shroom-Rho Kinase Complex Reveals a Binding Interface with Monomeric Shroom That Regulates Cell Morphology and Stimulates Kinase Activity.
abstract: Shroom-mediated remodeling of the actomyosin cytoskeleton is a critical driver of cellular shape and tissue morphology that underlies the development of many tissues including the neural tube, eye, intestines, and vasculature. Shroom uses a conserved SD2 domain to direct the subcellular localization of Rho-associated kinase (Rock), which in turn drives changes in the cytoskeleton and cellular morphology through its ability to phosphorylate and activate non-muscle myosin II. Here, we present the structure of the human Shroom-Rock binding module, revealing an unexpected stoichiometry for Shroom in which two Shroom SD2 domains bind independent surfaces on Rock. Mutation of interfacial residues impaired Shroom-Rock binding in vitro and resulted in altered remodeling of the cytoskeleton and loss of Shroom-mediated changes in cellular morphology. Additionally, we provide the first direct evidence that Shroom can function as a Rock activator. These data provide molecular insight into the Shroom-Rock interface and demonstrate that Shroom directly participates in regulating cytoskeletal dynamics, adding to its known role in Rock localization.
authors.affiliation: From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260.
authors.forename: Jenna K
authors.initials: JK
authors.lastname: Zalewski
authors.affiliation: From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260.
authors.forename: Joshua H
authors.initials: JH
authors.lastname: Mo
authors.affiliation: From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260.
authors.forename: Simone
authors.initials: S
authors.lastname: Heber
authors.affiliation: the Department of Biology, Brookhaven National Laboratory, Upton, New York 11973, and.
authors.forename: Annie
authors.initials: A
authors.lastname: Heroux
authors.affiliation: the Department of Pharmacology, University of Washington, Seattle, Washington 98195.
authors.forename: Richard G
authors.initials: RG
authors.lastname: Gardner
authors.affiliation: From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Hildebrand
authors.affiliation: From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, andyv@pitt.edu.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: VanDemark
mesh_terms: D051606:G-Protein-Coupled Receptor Kinase 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008840:Microfilament Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D024462:Myosin Type II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000072417:Protein Domains; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship
chemical_list: D008565:Membrane Proteins; D008840:Microfilament Proteins; D046912:Multiprotein Complexes; C539270:Shroom1 protein, human; D051606:G-Protein-Coupled Receptor Kinase 1; C496764:GRK1 protein, human; D024462:Myosin Type II
keywords: Ras homolog gene family, member A (RhoA); Rho-kinase; RhoA; Shroom; X-ray crystallography; coiled-coil; cytoskeleton; serine/threonine protein kinase; structural biology