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PMID: 21070952 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, N.I.H., Intramural Research Support, U.S. Gov't, Non-P.H.S.

Crystallographic and functional analysis of the ESCRT-I /HIV-1 Gag PTAP interaction.

Structure (London, England : 1993) ·Vol. 18 ·No. 11 ·2010-11-10 ·Pages 1536-47

Im YJ, Kuo L, Ren X, Burgos PV, Zhao XZ, Liu F, Burke TR, Bonifacino JS, Freed EO, Hurley JH

Abstract

Budding of HIV-1 requires the binding of the PTAP late domain of the Gag p6 protein to the UEV domain of the TSG101 subunit of ESCRT-I. The normal function of this motif in cells is in receptor downregulation. Here, we report the 1.4-1.6 Å structures of the human TSG101 UEV domain alone and with wild-type and mutant HIV-1 PTAP and Hrs PSAP nonapeptides. The hydroxyl of the Thr or Ser residue in the P(S/T)AP motif hydrogen bonds with the main chain of Asn69. Mutation of the Asn to Pro, blocking the main-chain amide, abrogates PTAP motif binding in vitro and blocks budding of HIV-1 from cells. N69P and other PTAP binding-deficient alleles of TSG101 did not rescue HIV-1 budding. However, the mutant alleles did rescue downregulation of endogenous EGF receptor. This demonstrates that the PSAP motif is not rate determining in EGF receptor downregulation under normal conditions.

MeSH Terms
Crystallography DNA-Binding Proteins/chemistry,isolation & purification,metabolism Endosomal Sorting Complexes Required for Transport/chemistry,isolation & purification,metabolism ErbB Receptors/metabolism HIV-1/chemistry HeLa Cells Humans Hydrogen Bonding Models, Molecular Molecular Dynamics Simulation Protein Conformation Protein Structure, Tertiary/genetics RNA Interference Transcription Factors/chemistry,isolation & purification,metabolism gag Gene Products, Human Immunodeficiency Virus/chemistry,isolation & purification,metabolism
Chemicals
DNA-Binding Proteins Endosomal Sorting Complexes Required for Transport Transcription Factors Tsg101 protein gag Gene Products, Human Immunodeficiency Virus ErbB Receptors
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Im Young Jun
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0580, USA.
Kuo Lillian
Ren Xuefeng
Burgos Patricia V
Zhao Xue Zhi
Liu Fa
Burke Terrence R
Bonifacino Juan S
Freed Eric O
Hurley James H
References (51)
51 references, click to expand
  1. The human endosomal sorting complex required for transport (ESCRT-I) and its role in HIV-1 budding.
    J Biol Chem. 2004 Aug 20;279(34):36059-71 PMID: 15218037
  2. Hrs interacts with sorting nexin 1 and regulates degradation of epidermal growth factor receptor.
    J Biol Chem. 2001 Mar 9;276(10):7069-78 PMID: 11110793
  3. MVB-12, a fourth subunit of metazoan ESCRT-I, functions in receptor downregulation.
    PLoS One. 2007 Sep 26;2(9):e956 PMID: 17895996
  4. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone.
    J Virol. 1986 Aug;59(2):284-91 PMID: 3016298
  5. Molecular mechanism of multivesicular body biogenesis by ESCRT complexes.
    Nature. 2010 Apr 8;464(7290):864-9 PMID: 20305637
  6. HIV Gag mimics the Tsg101-recruiting activity of the human Hrs protein.
    J Cell Biol. 2003 Aug 4;162(3):425-34 PMID: 12900394
  7. Ubiquitin recognition by the human TSG101 protein.
    Mol Cell. 2004 Mar 26;13(6):783-9 PMID: 15053872
  8. HIV-1 and Ebola virus encode small peptide motifs that recruit Tsg101 to sites of particle assembly to facilitate egress.
    Nat Med. 2001 Dec;7(12):1313-9 PMID: 11726971
  9. Effect of mutations affecting the p6 gag protein on human immunodeficiency virus particle release.
    Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3195-9 PMID: 2014240
  10. SAR by oxime-containing peptide libraries: application to Tsg101 ligand optimization.
    Chembiochem. 2008 Aug 11;9(12):2000-4 PMID: 18655064
  11. p6Gag is required for particle production from full-length human immunodeficiency virus type 1 molecular clones expressing protease.
    J Virol. 1995 Nov;69(11):6810-8 PMID: 7474093
  12. Proline-rich sequence recognition: II. Proteomics analysis of Tsg101 ubiquitin-E2-like variant (UEV) interactions.
    Mol Cell Proteomics. 2009 Nov;8(11):2474-86 PMID: 19542561
  13. Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding.
    Cell. 2001 Oct 5;107(1):55-65 PMID: 11595185
  14. Retrovirus budding.
    Annu Rev Cell Dev Biol. 2004;20:395-425 PMID: 15473846
  15. Structure of the Tsg101 UEV domain in complex with the PTAP motif of the HIV-1 p6 protein.
    Nat Struct Biol. 2002 Nov;9(11):812-7 PMID: 12379843
  16. Evidence for a functional interaction between the V1/V2 and C4 domains of human immunodeficiency virus type 1 envelope glycoprotein gp120.
    J Virol. 1994 Apr;68(4):2503-12 PMID: 8139032
  17. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
  18. HIV-1 drug resistance mutations: an updated framework for the second decade of HAART.
    AIDS Rev. 2008 Apr-Jun;10(2):67-84 PMID: 18615118
  19. Hrs regulates multivesicular body formation via ESCRT recruitment to endosomes.
    J Cell Biol. 2003 Aug 4;162(3):435-42 PMID: 12900395
  20. No strings attached: the ESCRT machinery in viral budding and cytokinesis.
    J Cell Sci. 2009 Jul 1;122(Pt 13):2167-77 PMID: 19535732
  21. Identification of human VPS37C, a component of endosomal sorting complex required for transport-I important for viral budding.
    J Biol Chem. 2005 Jan 7;280(1):628-36 PMID: 15509564
  22. Structural insights into endosomal sorting complex required for transport (ESCRT-I) recognition of ubiquitinated proteins.
    J Biol Chem. 2004 Jul 2;279(27):28689-96 PMID: 15044434
  23. Membrane scission by the ESCRT-III complex.
    Nature. 2009 Mar 12;458(7235):172-7 PMID: 19234443
  24. The late domain of human immunodeficiency virus type 1 p6 promotes virus release in a cell type-dependent manner.
    J Virol. 2002 Jan;76(1):105-17 PMID: 11739676
  25. Peptide inhibitors of HIV-1 egress.
    ACS Chem Biol. 2008 Dec 19;3(12):745-7 PMID: 19099395
  26. Membrane budding and scission by the ESCRT machinery: it's all in the neck.
    Nat Rev Mol Cell Biol. 2010 Aug;11(8):556-66 PMID: 20588296
  27. Identification of human MVB12 proteins as ESCRT-I subunits that function in HIV budding.
    Cell Host Microbe. 2007 Jul 12;2(1):41-53 PMID: 18005716
  28. TSG101 interaction with HRS mediates endosomal trafficking and receptor down-regulation.
    Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7626-31 PMID: 12802020
  29. Inhibition of HIV budding by a genetically selected cyclic peptide targeting the Gag-TSG101 interaction.
    ACS Chem Biol. 2008 Dec 19;3(12):757-64 PMID: 19053244
  30. The C-terminal portion of the Hrs protein interacts with Tsg101 and interferes with human immunodeficiency virus type 1 Gag particle production.
    J Virol. 2007 Mar;81(6):2909-22 PMID: 17182674
  31. Molecular architecture and functional model of the complete yeast ESCRT-I heterotetramer.
    Cell. 2007 May 4;129(3):485-98 PMID: 17442384
  32. Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors.
    Protein Expr Purif. 1999 Feb;15(1):34-9 PMID: 10024467
  33. Parallels between cytokinesis and retroviral budding: a role for the ESCRT machinery.
    Science. 2007 Jun 29;316(5833):1908-12 PMID: 17556548
  34. The ESCRT machinery in endosomal sorting of ubiquitylated membrane proteins.
    Nature. 2009 Mar 26;458(7237):445-52 PMID: 19325624
  35. Viral late domains.
    J Virol. 2002 May;76(10):4679-87 PMID: 11967285
  36. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  37. Mutations in erupted, the Drosophila ortholog of mammalian tumor susceptibility gene 101, elicit non-cell-autonomous overgrowth.
    Dev Cell. 2005 Nov;9(5):699-710 PMID: 16256744
  38. Biosynthesis, cleavage, and degradation of the human immunodeficiency virus 1 envelope glycoprotein gp160.
    Proc Natl Acad Sci U S A. 1988 Dec;85(24):9580-4 PMID: 2849111
  39. Structure of human TSG101 UEV domain.
    Acta Crystallogr D Biol Crystallogr. 2006 Apr;62(Pt 4):458-64 PMID: 16552148
  40. Overexpression of the N-terminal domain of TSG101 inhibits HIV-1 budding by blocking late domain function.
    Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):955-60 PMID: 11805336
  41. Mammalian class E vps proteins recognize ubiquitin and act in the removal of endosomal protein-ubiquitin conjugates.
    J Cell Biol. 2002 Apr 1;157(1):91-101 PMID: 11916981
  42. Mechanisms for enveloped virus budding: can some viruses do without an ESCRT?
    Virology. 2008 Mar 15;372(2):221-32 PMID: 18063004
  43. Mammalian tumor susceptibility gene 101 (TSG101) and the yeast homologue, Vps23p, both function in late endosomal trafficking.
    Traffic. 2000 Mar;1(3):248-58 PMID: 11208108
  44. Structure and functional interactions of the Tsg101 UEV domain.
    EMBO J. 2002 May 15;21(10):2397-406 PMID: 12006492
  45. Phaser crystallographic software.
    J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674 PMID: 19461840
  46. Tsg101, a homologue of ubiquitin-conjugating (E2) enzymes, binds the L domain in HIV type 1 Pr55(Gag).
    Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7724-9 PMID: 11427703
  47. Tsg101: a novel tumor susceptibility gene isolated by controlled homozygous functional knockout of allelic loci in mammalian cells.
    Cell. 1996 May 3;85(3):319-29 PMID: 8616888
  48. Molecular characterization of feline immunodeficiency virus budding.
    J Virol. 2008 Mar;82(5):2106-19 PMID: 18094166
  49. Response of a protein structure to cavity-creating mutations and its relation to the hydrophobic effect.
    Science. 1992 Jan 10;255(5041):178-83 PMID: 1553543
  50. Normalization of nomenclature for peptide motifs as ligands of modular protein domains.
    FEBS Lett. 2002 Feb 20;513(1):141-4 PMID: 11911894
  51. The cell biology of HIV-1 virion genesis.
    Cell Host Microbe. 2009 Jun 18;5(6):550-8 PMID: 19527882
Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
1878-4186
DOI
10.1016/j.str.2010.08.010
Published
2010-11-10
Pages
1536-47
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC3124085
Subset
IM
Grants
Intramural NIH HHS · ZIA DK036125-04 · United States
NCI NIH HHS · Y1-CO-1020 · United States
NIGMS NIH HHS · Y1-GM-1104 · United States
External Links
PubMed source DOI: 10.1016/j.str.2010.08.010 PMC Full Text View journal details
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