title: Key interactions for clathrin coat stability.
abstract: Clathrin-coated vesicles are major carriers of vesicular traffic in eukaryotic cells. This endocytic pathway relies on cycles of clathrin coat assembly and Hsc70-mediated disassembly. Here we identify histidine residues as major determinants of lattice assembly and stability. They are located at the invariant interface between the proximal and distal segments of clathrin heavy chains, in triskelions centered on two adjacent vertices of the coated-vesicle lattice. Mutation of these histidine residues to glutamine alters the pH dependence of coat stability. We then describe single-particle fluorescence imaging experiments in which we follow the effect of these histidine mutations on susceptibility to Hsc70-dependent uncoating. Coats destabilized by these mutations require fewer Hsc70 molecules to initiate disassembly, as predicted by a model in which Hsc70 traps conformational distortions during the auxilin- and Hsc70:ATP-mediated uncoating reaction.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Centre for Vascular Research, University of New South Wales, Sydney NSW 2052 Australia. Electronic address: till.boecking@unsw.edu.au.
authors.forename: Till
authors.initials: T
authors.lastname: Böcking
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: François
authors.initials: F
authors.lastname: Aguet
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Program in Cellular and Molecular Medicine at Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Iris
authors.initials: I
authors.lastname: Rapoport
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Program in Cellular and Molecular Medicine at Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Manuel
authors.initials: M
authors.lastname: Banzhaf
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Program in Cellular and Molecular Medicine at Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Anan
authors.initials: A
authors.lastname: Yu
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Program in Cellular and Molecular Medicine at Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Jean Christophe
authors.initials: JC
authors.lastname: Zeeh
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Program in Cellular and Molecular Medicine at Boston Children's Hospital, Boston, MA 02115, USA. Electronic address: kirchhausen@crystal.harvard.edu.
authors.forename: Tom
authors.initials: T
authors.lastname: Kirchhausen
mesh_terms: D000818:Animals; D034021:Auxilins / Q000737:chemistry; D001665:Binding Sites; D002417:Cattle; D033922:Clathrin Heavy Chains / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D033941:Clathrin Light Chains / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D022163:Clathrin-Coated Vesicles / Q000737:chemistry / Q000648:ultrastructure; D050883:HSC70 Heat-Shock Proteins / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry; D009154:Mutation; D055550:Protein Stability; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure
chemical_list: D034021:Auxilins; D033941:Clathrin Light Chains; D050883:HSC70 Heat-Shock Proteins; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; D033922:Clathrin Heavy Chains; D006639:Histidine