title: A dynamin mutant defines a superconstricted prefission state.
abstract: Dynamin is a 100 kDa GTPase that organizes into helical assemblies at the base of nascent clathrin-coated vesicles. Formation of these oligomers stimulates the intrinsic GTPase activity of dynamin, which is necessary for efficient membrane fission during endocytosis. Recent evidence suggests that the transition state of dynamin's GTP hydrolysis reaction serves as a key determinant of productive fission. Here, we present the structure of a transition-state-defective dynamin mutant K44A trapped in a prefission state at 12.5 Å resolution. This structure constricts to 3.7 nm, reaching the theoretical limit required for spontaneous membrane fission. Computational docking indicates that the ground-state conformation of the dynamin polymer is sufficient to achieve this superconstricted prefission state and reveals how a two-start helical symmetry promotes the most efficient packing of dynamin tetramers around the membrane neck. These data suggest a model for the assembly and regulation of the minimal dynamin fission machine.
authors.affiliation: Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Anna C
authors.initials: AC
authors.lastname: Sundborger
authors.affiliation: Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Shunming
authors.initials: S
authors.lastname: Fang
authors.affiliation: Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Jürgen A
authors.initials: JA
authors.lastname: Heymann
authors.affiliation: Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Pampa
authors.initials: P
authors.lastname: Ray
authors.affiliation: Department of Molecular Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850, USA. Electronic address: chappie@cornell.edu.
authors.forename: Joshua S
authors.initials: JS
authors.lastname: Chappie
authors.affiliation: Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: jennyh@helix.nih.gov.
authors.forename: Jenny E
authors.initials: JE
authors.lastname: Hinshaw
mesh_terms: D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000737:chemistry / Q000378:metabolism; D034281:Dynamins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009154:Mutation; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary
chemical_list: D006160:Guanosine Triphosphate; D034281:Dynamins
references.citation: J Struct Biol. 2008 Aug;163(2):163-74
references.pmid: 18572416
references.citation: Proc Natl Acad Sci U S A. 2013 Nov 5;110(45):18037-41
references.pmid: 24106306
references.citation: Cell. 2012 Mar 30;149(1):137-45
references.pmid: 22464326
references.citation: Biophys J. 2003 Jul;85(1):85-96
references.pmid: 12829467
references.citation: Nat Struct Mol Biol. 2004 Jun;11(6):574-5
references.pmid: 15133500
references.citation: J Neurochem. 2007 Aug;102(3):931-43
references.pmid: 17437541
references.citation: Nat Rev Mol Cell Biol. 2012 Jan 11;13(2):75-88
references.pmid: 22233676
references.citation: Nature. 2011 Sep 18;477(7366):561-6
references.pmid: 21927001
references.citation: Science. 2013 Mar 22;339(6126):1433-6
references.pmid: 23520112
references.citation: Nat Struct Mol Biol. 2011 Jan;18(1):20-6
references.pmid: 21170049
references.citation: Cell. 2008 Dec 26;135(7):1263-75
references.pmid: 19084268
references.citation: J Struct Biol. 2007 Jan;157(1):3-18
references.pmid: 17011211
references.citation: Nat Cell Biol. 2001 Oct;3(10):922-6
references.pmid: 11584275
references.citation: Mol Biol Cell. 2009 Nov;20(22):4630-9
references.pmid: 19776347
references.citation: EMBO J. 2008 Jan 9;27(1):27-37
references.pmid: 18079695
references.citation: J Struct Biol. 2004 Sep;147(3):259-67
references.pmid: 15450295
references.citation: Cell. 2012 Oct 26;151(3):619-29
references.pmid: 23101629
references.citation: J Struct Biol. 2007 Jan;157(1):83-94
references.pmid: 16919474
references.citation: Dev Cell. 2010 May 18;18(5):687-8
references.pmid: 20493800
references.citation: Cell. 1994 Oct 21;79(2):199-209
references.pmid: 7954789
references.citation: Cell. 2011 Sep 30;147(1):209-22
references.pmid: 21962517
references.citation: Nature. 2011 Sep 18;477(7366):556-60
references.pmid: 21927000
references.citation: J Cell Biol. 1994 Nov;127(4):915-34
references.pmid: 7962076
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4141-6
references.pmid: 20160074
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13359-64
references.pmid: 19666604
references.citation: PLoS One. 2013;8(1):e55691
references.pmid: 23383266
references.citation: Annu Rev Cell Dev Biol. 2011;27:79-105
references.pmid: 21599493
references.citation: Mol Biol Cell. 2009 Aug;20(15):3561-71
references.pmid: 19515832
references.citation: J Cell Sci. 2011 Jan 1;124(Pt 1):133-43
references.pmid: 21172823
references.citation: Nature. 2006 May 25;441(7092):528-31
references.pmid: 16648839
references.citation: J Biol Chem. 1996 Sep 13;271(37):22310-4
references.pmid: 8798389
references.citation: J Cell Sci. 2009 Oct 1;122(Pt 19):3427-31
references.pmid: 19759282
references.citation: Cell. 2008 Dec 26;135(7):1276-86
references.pmid: 19084269
references.citation: Cell. 1998 Jun 12;93(6):1021-9
references.pmid: 9635431
references.citation: Biochemistry. 2000 Oct 10;39(40):12485-93
references.pmid: 11015230
references.citation: Structure. 2007 Oct;15(10):1190-202
references.pmid: 17937909
references.citation: J Cell Sci. 2013 Jul 1;126(Pt 13):2773-84
references.pmid: 23781021
references.citation: Mol Biol Cell. 2014 Mar;25(6):879-90
references.pmid: 24478459
references.citation: Nature. 2010 May 27;465(7297):435-40
references.pmid: 20428113