CoChat
Sign Up
Cited by 133HIV Research and TreatmentAdvanced Electron Microscopy Techniques and ApplicationsT-cell and Retrovirus Studies

Cryo Electron Tomography of Native HIV-1 Budding Sites

Open access
Carlson, Lars-Anders, de Marco, Alex, Oberwinkler, Heike, Habermann, Anja, Briggs, John A. G., Kräusslich, Hans-Georg, Grünewald, Kay·2010·PLoS Pathogens
DOI 10.1371/journal.ppat.1001173PMC PMC2991257
Open this paper in CoChat — free accountOpen PDF

Abstract

The structure of immature and mature HIV-1 particles has been analyzed in detail by cryo electron microscopy, while no such studies have been reported for cellular HIV-1 budding sites. Here, we established a system for studying HIV-1 virus-like particle assembly and release by cryo electron tomography of intact human cells. The lattice of the structural Gag protein in budding sites was indistinguishable from that of the released immature virion, suggesting that its organization is determined at the assembly site without major subsequent rearrangements. Besides the immature lattice, a previously not described Gag lattice was detected in some budding sites and released particles; this lattice was found at high frequencies in a subset of infected T-cells. It displays the same hexagonal symmetry and spacing in the MA-CA layer as the immature lattice, but lacks density corresponding to NC-RNA-p6. Buds and released particles carrying this lattice consistently lacked the viral ribonucleoprotein complex, suggesting that they correspond to aberrant products due to premature proteolytic activation. We hypothesize that cellular and/or viral factors normally control the onset of proteolytic maturation during assembly and release, and that this control has been lost in a subset of infected T-cells leading to formation of aberrant particles.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.ppat.1001173

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Premature proteolytic cleavage can apparently occur in a subset of cells infected with adenoviral vectors expressing HIV-1 Gag and Gag-Pol, leading to cleavage between CA and NC before budding, and preventing RNP incorporation into particles.

    “Premature proteolytic cleavage can apparently occur in a subset of cells infected with adenoviral vectors expressing HIV-1 Gag and Gag-Pol, leading to cleavage between CA and NC before budding, and preventing RNP incorporation into particles”

    From Retrovirus maturation-an extraordinary structural transformation (2016) · cites this paper as [62]

    Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 38

  1. [1]Freed EO 1998 HIV-1 gag proteins: diverse functions in the virus life cycle. Virology 251 1 15 9813197
  2. [2]Hurley JH 2008 ESCRT complexes and the biogenesis of multivesicular bodies. Curr Opin Cell Biol 20 4 11 18222686 PMC2282067
  3. [3]Morita E Sundquist WI 2004 Retrovirus budding. Ann Rev Cell Dev Biol 20 395 425 15473846
  4. [4]Williams RL Urbe S 2007 The emerging shape of the ESCRT machinery. Nat Rev Mol Cell Biol 8 355 368 17450176
  5. [5]Briggs JAG Simon MN Gross I Kräusslich HG Fuller SD 2004 The stoichiometry of Gag protein in HIV-1. Nat Struct Mol Biol 11 672 675 15208690
  6. [6]Garrus JE von Schwedler UK Pornillos OW Morham SG Zavitz KH 2001 Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding. Cell 107 55 65 11595185
  7. [7]VerPlank L Bouamr F LaGrassa TJ Agresta B Kikonyogo A 2001 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 98 7724 7729 11427703 PMC35409
  8. [8]Coffin JM Hughes SH Varmus HE 1997 Retroviruses: Cold Spring Harbor Laboratory Press. 21433340
  9. [9]Pettit SC Moody MD Wehbie RS Kaplan AH Nantermet PV 1994 The p2 domain of human immunodeficiency virus type 1 Gag regulates sequential proteolytic processing and is required to produce fully infectious virions. J Virol 68 8017 8027 7966591 PMC237265
  10. [11]Coren LV Thomas JA Chertova E Sowder RC Gagliardi TD 2007 Mutational analysis of the C-terminal Gag cleavage sites in human immunodeficiency virus type 1. J Virol 81 10047 10054 17634233 PMC2045408
  11. [12]Checkley MA Luttge BG Soheilian F Nagashima K Freed EO 2010 The capsid-spacer peptide 1 Gag processing intermediate is a dominant-negative inhibitor of HIV-1 maturation. Virology 400 137 144 20172577 PMC2838455
  12. [13]Lee SK Harris J Swanstrom R 2009 A strongly transdominant mutation in the human immunodeficiency virus type 1 gag gene defines an Achilles heel in the virus life cycle. J Virol 83 8536 8543 19515760 PMC2738153
  13. [14]Müller B Anders M Akiyama H Welsch S Glass B 2009 HIV-1 Gag processing intermediates trans-dominantly interfere with HIV-1 infectivity. J Biol Chem 284 29692 29703 19666477 PMC2785601
  14. [15]Kräusslich HG 1991 Human immunodeficiency virus proteinase dimer as component of the viral polyprotein prevents particle assembly and viral infectivity. Proc Natl Acad Sci U S A 88 3213 3217 2014242 PMC51416
  15. [16]Kaplan AH Zack JA Knigge M Paul DA Kempf DJ 1993 Partial inhibition of the human immunodeficiency virus type 1 protease results in aberrant virus assembly and the formation of noninfectious particles. J Virol 67 4050 4055 8510215 PMC237772
  16. [17]Moore MD Fu W Soheilian F Nagashima K Ptak RG 2008 Suboptimal inhibition of protease activity in human immunodeficiency virus type 1: effects on virion morphogenesis and RNA maturation. Virology 379 152 160 18657842 PMC2577075
  17. [18]Benjamin J Ganser-Pornillos BK Tivol WF Sundquist WI Jensen GJ 2005 Three-dimensional structure of HIV-1 virus-like particles by electron cryotomography. J Mol Biol 346 577 588 15670606 PMC6608732
  18. [20]Briggs JAG Grünewald K Glass B Förster F Kräusslich HG 2006 The mechanism of HIV-1 core assembly: Insights from three-dimensional reconstructions of authentic virions. Structure 14 15 20 16407061
  19. [21]Liu J Bartesaghi A Borgnia MJ Sapiro G Subramaniam S 2008 Molecular architecture of native HIV-1 gp120 trimers. Nature 455 109 113 18668044 PMC2610422
  20. [22]Wright ER Schooler JB Ding HJ Kieffer C Fillmore C 2007 Electron cryotomography of immature HIV-1 virions reveals the structure of the CA and SP1 Gag shells. EMBO J 26 2218 2226 17396149 PMC1852790
  21. [23]Zhu P Liu J Bess J Jr Chertova E Lifson JD 2006 Distribution and three-dimensional structure of AIDS virus envelope spikes. Nature 441 847 852 16728975
  22. [24]Zhu P Winkler H Chertova E Taylor KA Roux KH 2008 Cryoelectron tomography of HIV-1 envelope spikes: further evidence for tripod-like legs. PLoS Pathog 4 e1000203 19008954 PMC2577619
  23. [25]Briggs JAG Wilk T Welker R Kräusslich HG Fuller SD 2003 Structural organization of authentic, mature HIV-1 virions and cores. EMBO J 22 1707 1715 12660176 PMC152888
  24. [27]Li S Hill CP Sundquist WI Finch JT 2000 Image reconstructions of helical assemblies of the HIV-1CA protein. Nature 407 409 413 11014200
  25. [28]Ganser BK Li S Klishko VY Finch JT Sundquist WI 1999 Assembly and analysis of conical models for the HIV-1 core. Science 283 80 83 9872746
  26. [29]Wagner R Graf M Bieler K Wolf H Grunwald T 2000 Rev-independent expression of synthetic gag-pol genes of human immunodeficiency virus type 1 and simian immunodeficiency virus: implications for the safety of lentiviral vectors. Hum Gene Ther 11 2403 2413 11096444
  27. [30]Medalia O Weber I Frangakis AS Nicastro D Gerisch G 2002 Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 298 1209 1213 12424373
  28. [31]Ott DE Coren LV Kane BP Busch LK Johnson DG 1996 Cytoskeletal proteins inside human immunodeficiency virus type 1 virions. J Virol 70 7734 7743 8892894 PMC190843
  29. [32]Gladnikoff M Shimoni E Gov NS Rousso I 2009 Retroviral assembly and budding occur through an actin-driven mechanism. Biophys J 97 2419 2428 19883584 PMC2770610
  30. [33]Crist RM Datta SA Stephen AG Soheilian F Mirro J 2009 Assembly properties of human immunodeficiency virus type 1 Gag-leucine zipper chimeras: implications for retrovirus assembly. J Virol 83 2216 2225 19073719 PMC2643709
  31. [34]Hockley DJ Nermut MV Grief C Jowett JB Jones IM 1994 Comparative morphology of Gag protein structures produced by mutants of the gag gene of human immunodeficiency virus type 1. J Gen Virol 75 Pt 11 2985 2997 7964608
  32. [35]Ott DE Coren LV Chertova EN Gagliardi TD Nagashima K 2003 Elimination of protease activity restores efficient virion production to a human immunodeficiency virus type 1 nucleocapsid deletion mutant. J Virol 77 5547 5556 12719547 PMC154014
  33. [36]Popova E Popov S Gottlinger HG 2010 Human Immunodeficiency Virus Type 1 Nucleocapsid p1 Confers ESCRT Pathway Dependence. J Virol 84 6590 6597 20427536 PMC2903293
  34. [37]Mastronarde DN 2005 Automated electron microscope tomography using robust prediction of specimen movements. J Struct Biol 152 36 51 16182563
  35. [38]Frank J 2006 Electron Tomography: Methods for Three-Dimensional Visualization of Structures in the Cell Berlin: Springer
  36. [39]Nickell S Förster F Linaroudis A Del Net W Beek F 2005 TOM software toolbox: acquisition and analysis for electron tomography. Journal of Structural Biology 149 227 234 15721576
  37. [40]Kremer JR Mastronarde DN McIntosh JR 1996 Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116 71 76 8742726
  38. [41]Westfall PH Young SS 1993 Resampling-based multiple testing: examples and methods for P-value adjustment: Wiley-Interscience 360

Cited by

Sorted by citations

Authors
Carlson, L.-A., de Marco, A., Oberwinkler, H., Habermann, A., Briggs, J. A. G., Kräusslich, H.-G., & Grünewald, K.
Affiliations
  • University Hospital Heidelberg (DE)
  • European Molecular Biology Laboratory (DE)
  • Centre for Human Genetics (GB)
Corresponding author
  • Kräusslich, Hans-Georg
  • Grünewald, Kay
ORCID
Venue
PLoS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2010-11-24
Volume
6
Issue
11
Pages
e1001173
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1001173
PMC
PMC2991257
OpenAlex
W1983749753
Cited by
133
References
3
Citations by year
2026: 2 · 2025: 2 · 2024: 6 · 2023: 7 · 2022: 9 · 2021: 7 · 2020: 7 · 2019: 5 · 2018: 13 · 2017: 6 · 2016: 8 · 2015: 8 · 2014: 15 · 2013: 12 · 2012: 10
FWCI
4.17
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Max Planck Institute for Plasma Physics (submittedVersion)
  • Figshare (submittedVersion, cc-by)
  • MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
Funders
  • Wellcome Trust (090895)
  • Deutsche Forschungsgemeinschaft (SP1175, SFB 638)
Topics
  • Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and Retrovirus Studies
Keywords
virus-like particle assembly, immature virions, viral ribonucleoprotein, HIV-1 structural proteins
MeSH terms
Cells, Cultured, Glioblastoma, Humans, RNA, Viral, T-Lymphocytes, Virion, HIV-1, Cryoelectron Microscopy, gag Gene Products, Human Immunodeficiency Virus, Electron Microscope Tomography

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review