title: Binding of Substrates to the Central Pore of the Vps4 ATPase Is Autoinhibited by the Microtubule Interacting and Trafficking (MIT) Domain and Activated by MIT Interacting Motifs (MIMs).
abstract: The endosomal sorting complexes required for transport (ESCRT) pathway drives reverse topology membrane fission events within multiple cellular pathways, including cytokinesis, multivesicular body biogenesis, repair of the plasma membrane, nuclear membrane vesicle formation, and HIV budding. The AAA ATPase Vps4 is recruited to membrane necks shortly before fission, where it catalyzes disassembly of the ESCRT-III lattice. The N-terminal Vps4 microtubule-interacting and trafficking (MIT) domains initially bind the C-terminal MIT-interacting motifs (MIMs) of ESCRT-III subunits, but it is unclear how the enzyme then remodels these substrates in response to ATP hydrolysis. Here, we report quantitative binding studies that demonstrate that residues from helix 5 of the Vps2p subunit of ESCRT-III bind to the central pore of an asymmetric Vps4p hexamer in a manner that is dependent upon the presence of flexible nucleotide analogs that can mimic multiple states in the ATP hydrolysis cycle. We also find that substrate engagement is autoinhibited by the Vps4p MIT domain and that this inhibition is relieved by binding of either Type 1 or Type 2 MIM elements, which bind the Vps4p MIT domain through different interfaces. These observations support the model that Vps4 substrates are initially recruited by an MIM-MIT interaction that activates the Vps4 central pore to engage substrates and generate force, thereby triggering ESCRT-III disassembly.
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650.
authors.forename: Han
authors.initials: H
authors.lastname: Han
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650.
authors.forename: Nicole
authors.initials: N
authors.lastname: Monroe
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650.
authors.forename: Jörg
authors.initials: J
authors.lastname: Votteler
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650.
authors.forename: Binita
authors.initials: B
authors.lastname: Shakya
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650 wes@biochem.utah.edu.
authors.forename: Wesley I
authors.initials: WI
authors.lastname: Sundquist
authors.affiliation: From the Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112-5650 chris@biochem.utah.edu.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Hill
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry / Q000378:metabolism*; D011992:Endosomes; D005454:Fluorescence Polarization; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D006868:Hydrolysis; D008870:Microtubules / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: C524222:DID4 protein, S cerevisiae; D056827:Endosomal Sorting Complexes Required for Transport; D029701:Saccharomyces cerevisiae Proteins; C107528:VPS4 protein, S cerevisiae; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases
keywords: ATPases associated with diverse cellular activities (AAA); endosomal sorting complexes required for transport (ESCRT); enzyme mechanism; fluorescence anisotropy; protein-protein interaction
references.citation: J Biol Chem. 2002 Sep 20;277(38):34760-5
references.pmid: 12080075
references.citation: J Cell Sci. 2003 Oct 1;116(Pt 19):3957-70
references.pmid: 12953057
references.citation: EMBO J. 2004 Mar 10;23(5):1030-9
references.pmid: 14988733
references.citation: EMBO J. 2004 Jul 21;23(14):2734-44
references.pmid: 15229653
references.citation: Nature. 1994 Aug 25;370(6491):621-8
references.pmid: 8065448
references.citation: EMBO J. 1998 Jun 1;17(11):2982-93
references.pmid: 9606181
references.citation: J Biol Chem. 2005 Mar 18;280(11):10548-55
references.pmid: 15644320
references.citation: Nat Rev Mol Cell Biol. 2005 Jul;6(7):519-29
references.pmid: 16072036
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13813-8
references.pmid: 16174732
references.citation: EMBO J. 2005 Oct 19;24(20):3658-69
references.pmid: 16193069
references.citation: J Cell Biol. 2006 Feb 27;172(5):705-17
references.pmid: 16505166
references.citation: Proc Natl Acad Sci U S A. 2006 Apr 18;103(16):6202-7
references.pmid: 16601096
references.citation: Annu Rev Biophys Biomol Struct. 2006;35:93-114
references.pmid: 16689629
references.citation: Dev Cell. 2006 Jun;10(6):821-30
references.pmid: 16740483
references.citation: Nature. 2006 Jul 20;442(7100):270-5
references.pmid: 16855583
references.citation: J Cell Biol. 2007 Mar 26;176(7):995-1005
references.pmid: 17389232
references.citation: Science. 2007 Jun 29;316(5833):1908-12
references.pmid: 17556548
references.citation: EMBO J. 2007 Oct 3;26(19):4215-27
references.pmid: 17853893
references.citation: Nature. 2007 Oct 11;449(7163):735-9
references.pmid: 17928861
references.citation: Nature. 2007 Oct 11;449(7163):740-4
references.pmid: 17928862
references.citation: J Mol Biol. 2007 Nov 30;374(3):655-70
references.pmid: 17949747
references.citation: Dev Cell. 2008 Jan;14(1):50-61
references.pmid: 18194652
references.citation: Nature. 2008 Jan 17;451(7176):363-7
references.pmid: 18202664
references.citation: J Cell Biol. 2008 Jan 28;180(2):389-402
references.pmid: 18209100
references.citation: Mol Biol Cell. 2008 Feb;19(2):465-74
references.pmid: 18032582
references.citation: J Mol Biol. 2008 Mar 21;377(2):352-63
references.pmid: 18272179
references.citation: J Mol Biol. 2013 Jan 23;425(2):199-213
references.pmid: 23147216
references.citation: Annu Rev Biochem. 2013;82:663-92
references.pmid: 23527693
references.citation: Structure. 2013 Sep 3;21(9):1551-62
references.pmid: 24010714
references.citation: Cell Host Microbe. 2013 Sep 11;14(3):232-41
references.pmid: 24034610
references.citation: J Biol Chem. 2013 Sep 13;288(37):26810-9
references.pmid: 23913684
references.citation: Genes Dev. 2013 Nov 15;27(22):2500-11
references.pmid: 24240239
references.citation: J Mol Biol. 2014 Feb 6;426(3):510-25
references.pmid: 24161953
references.citation: J Mol Biol. 2008 Mar 21;377(2):364-77
references.pmid: 18280501
references.citation: Mol Biol Cell. 2008 Jun;19(6):2661-72
references.pmid: 18385515
references.citation: Dev Cell. 2008 Jul;15(1):62-73
references.pmid: 18606141
references.citation: Science. 2008 Sep 5;321(5894):1354-7
references.pmid: 18687924
references.citation: Nat Struct Mol Biol. 2008 Nov;15(11):1147-51
references.pmid: 18931677
references.citation: J Mol Biol. 2008 Dec 26;384(4):878-95
references.pmid: 18929572
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4846-51
references.pmid: 21383202
references.citation: Science. 2011 Mar 25;331(6024):1616-20
references.pmid: 21310966
references.citation: Nat Cell Biol. 2011 Apr;13(4):469-74
references.pmid: 21394086
references.citation: J Struct Funct Genomics. 2011 Jul;12(2):55-62
references.pmid: 21360289
references.citation: Dev Cell. 2011 Jul 19;21(1):77-91
references.pmid: 21763610
references.citation: Science. 2008 Dec 12;322(5908):1710-3
references.pmid: 19008417
references.citation: Structure. 2009 Mar 11;17(3):427-37
references.pmid: 19278657
references.citation: Traffic. 2008 Dec;9(12):2180-9
references.pmid: 18796009
references.citation: Cell Host Microbe. 2009 Jun 18;5(6):550-8
references.pmid: 19527882
references.citation: Nat Struct Mol Biol. 2009 Jul;16(7):754-62
references.pmid: 19525971
references.citation: Mol Biol Cell. 2009 Aug;20(15):3514-24
references.pmid: 19477918
references.citation: Cell. 2009 Oct 30;139(3):523-34
references.pmid: 19879839
references.citation: Proteomics. 2009 Nov;9(21):4997-5000
references.pmid: 19810033
references.citation: Mol Biol Cell. 2010 Mar 15;21(6):1059-71
references.pmid: 20110351
references.citation: Nat Rev Mol Cell Biol. 2010 Aug;11(8):556-66
references.pmid: 20588296
references.citation: Structure. 2010 Aug 11;18(8):976-84
references.pmid: 20696398
references.citation: J Biol Chem. 2010 Nov 12;285(46):35428-38
references.pmid: 20805225
references.citation: J Mol Biol. 2011 Jul 22;410(4):525-33
references.pmid: 21762798
references.citation: Curr Biol. 2011 Dec 6;21(23):1951-9
references.pmid: 22100064
references.citation: Biochim Biophys Acta. 2012 Jan;1823(1):172-81
references.pmid: 21925211
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4146-51
references.pmid: 22315426
references.citation: Nat Cell Biol. 2012 Jul;14(7):677-85
references.pmid: 22660413
references.citation: Annu Rev Cell Dev Biol. 2012;28:337-62
references.pmid: 22831642
references.citation: Science. 2014 Feb 28;343(6174):1247136
references.pmid: 24482116
references.citation: J Biol Chem. 2014 Apr 11;289(15):10378-86
references.pmid: 24567329
references.citation: Cell. 2014 Oct 9;159(2):388-401
references.pmid: 25303532
references.citation: Nature. 2015 Feb 5;518(7537):61-7
references.pmid: 25581794
references.citation: Structure. 2000 Jun 15;8(6):567-73
references.pmid: 10873854
references.citation: J Immunol. 2001 Jun 15;166(12):7309-18
references.pmid: 11390481
references.citation: Genes Dev. 2001 Sep 1;15(17):2282-94
references.pmid: 11544185
references.citation: Nat Cell Biol. 2002 May;4(5):E113-6
references.pmid: 11988749