CoChat
Sign Up
Cited by 73HIV Research and TreatmentLipid Membrane Structure and BehaviorLipid metabolism and biosynthesis

Quantification of phosphoinositides reveals strong enrichment of PIP2 in HIV-1 compared to producer cell membranes

Open access
Mücksch, Frauke, Citir, Mevlut, Lüchtenborg, Christian, Glass, Bärbel, Traynor-Kaplan, Alexis, Schultz, Carsten, Brügger, Britta, Kräusslich, Hans-Georg·2019·Scientific Reports
DOI 10.1038/s41598-019-53939-zPMC PMC6881329
Open this paper in CoChat — free accountOpen PDF

Abstract

Human immunodeficiency virus type 1 (HIV-1) acquires its lipid envelope during budding from the plasma membrane of the host cell. Various studies indicated that HIV-1 membranes differ from producer cell plasma membranes, suggesting budding from specialized membrane microdomains. The phosphoinositide PI(4,5)P 2 has been of particular interest since PI(4,5)P 2 is needed to recruit the viral structural polyprotein Gag to the plasma membrane and thus facilitates viral morphogenesis. While there is evidence for an enrichment of PIP 2 in HIV-1, fully quantitative analysis of all phosphoinositides remains technically challenging and therefore has not been reported, yet. Here, we present a comprehensive analysis of the lipid content of HIV-1 and of plasma membranes from infected and non-infected producer cells, resulting in a total of 478 quantified lipid compounds, including molecular species distribution of 25 different lipid classes. Quantitative analyses of phosphoinositides revealed strong enrichment of PIP 2 , but also of PIP 3 , in the viral compared to the producer cell plasma membrane. We calculated an average of ca. 8,000 PIP 2 molecules per HIV-1 particle, three times more than Gag. We speculate that the high density of PIP 2 at the HIV-1 assembly site is mediated by transient interactions with viral Gag polyproteins, facilitating PIP 2 concentration in this microdomain. These results are consistent with our previous observation that PIP 2 is not only required for recruiting, but also for stably maintaining Gag at the plasma membrane. We believe that this quantitative analysis of the molecular anatomy of the HIV-1 lipid envelope may serve as standard reference for future investigations.

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 · 10.1038/s41598-019-53939-z

Cites

6 of 6 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?

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: 48

  1. [1]Waheed AA Freed EO Lipids and membrane microdomains in HIV-1 replication Virus Res. 2009 143 162 76 10.1016/j.virusres.2009.04.007 19383519 PMC2731011 · DOI
  2. [2]Sundquist WI Kräusslich H-G HIV-1 assembly, budding, and maturation Cold Spring Harb. Perspect. Med. 2012 2 a006924 10.1101/cshperspect.a006924 22762019 PMC3385941 · DOI · Paper page
  3. [3]Freed EO HIV-1 assembly, release and maturation Nat. Rev. Microbiol. 2015 13 484 496 10.1038/nrmicro3490 26119571 PMC6936268 · DOI · Paper page
  4. [4]Aloia RC Jensen FC Curtain CC Mobley PW Gordon LM Lipid composition and fluidity of the human immunodeficiency virus Proc. Natl. Acad. Sci. USA 1988 85 900 4 10.1073/pnas.85.3.900 2829209 PMC279664 · DOI
  5. [5]Aloia RC Tian H Jensen FC Lipid composition and fluidity of the human immunodeficiency virus envelope and host cell plasma membranes Proc. Natl. Acad. Sci. 1993 90 5181 5185 10.1073/pnas.90.11.5181 8389472 PMC46679 · DOI
  6. [6]Nguyen DH Hildreth JEK Evidence for Budding of Human Immunodeficiency Virus Type 1 Selectively from Glycolipid-Enriched Membrane Lipid Rafts J. Virol. 2000 74 3264 3272 10.1128/JVI.74.7.3264-3272.2000 10708443 PMC111827 · DOI
  7. [7]Lindwasser OW Resh MD Human immunodeficiency virus type 1 Gag contains a dileucine-like motif that regulates association with multivesicular bodies J. Virol. 2004 78 6013 6023 10.1128/JVI.78.11.6013-6023.2004 15140999 PMC415809 · DOI
  8. [8]Ono A Freed EO Plasma membrane rafts play a critical role in HIV-1 assembly and release Proc. Natl. Acad. Sci. USA 2001 98 13925 30 10.1073/pnas.241320298 11717449 PMC61143 · DOI
  9. [9]Bhattacharya J Repik A Clapham PR Gag regulates association of human immunodeficiency virus type 1 envelope with detergent-resistant membranes J. Virol. 2006 80 5292 300 10.1128/JVI.01469-05 16699009 PMC1472128 · DOI
  10. [10]Brügger B The HIV lipidome A raft with an unusual composition Proc. Natl. Acad. Sci. USA 2006 103 2641 2646 10.1073/pnas.0511136103 16481622 PMC1413831 · DOI
  11. [12]Lorizate M Comparative lipidomics analysis of HIV-1 particles and their producer cell membrane in different cell lines Cell. Microbiol. 2013 15 292 304 10.1111/cmi.12101 23279151 · DOI
  12. [13]Lorizate M Probing HIV-1 membrane liquid order by Laurdan staining reveals producer cell-dependent differences J. Biol. Chem. 2009 284 22238 22247 10.1074/jbc.M109.029256 19553682 PMC2755948 · DOI
  13. [14]Manes S Membrane raft microdomains mediate lateral assemblies required for HIV-1 infection EMBO Rep. 2000 1 190 196 10.1093/embo-reports/kvd025 11265761 PMC1084257 · DOI
  14. [15]Liao Z Graham DR Hildreth JEK Lipid rafts and HIV pathogenesis: virion-associated cholesterol is required for fusion and infection of susceptible cells AIDS Res. Hum. Retroviruses 2003 19 675 687 10.1089/088922203322280900 13678470 · DOI
  15. [16]Graham DRM Chertova E Hilburn JM Arthur LO Hildreth JEK Cholesterol Depletion of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus with -Cyclodextrin Inactivates and Permeabilizes the Virions: Evidence for Virion-Associated Lipid Rafts J. Virol. 2003 77 8237 8248 10.1128/JVI.77.15.8237-8248.2003 12857892 PMC165256 · DOI
  16. [17]Waheed AA Inhibition of HIV-1 replication by amphotericin B methyl ester: selection for resistant variants J. Biol. Chem. 2006 281 28699 28711 10.1074/jbc.M603609200 16882663 · DOI
  17. [18]Nieto-Garai JA Lipidomimetic Compounds Act as HIV-1 Entry Inhibitors by Altering Viral Membrane Structure Front. Immunol. 2018 9 1983 10.3389/fimmu.2018.01983 30233582 PMC6131562 · DOI
  18. [20]Freed E Orenstein JANM Buckler-white AJ Martin MA Single Amino Acid Changes in the Human Immunodeficiency Virus Type 1 Matrix Protein Block Virus Particle Production J. Virol. 1994 68 5311 5320 8035531 PMC236481
  19. [21]Ono A Huang M Freed EO Characterization of human immunodeficiency virus type 1 matrix revertants: effects on virus assembly, Gag processing, and Env incorporation into virions J. Virol. 1997 71 4409 18 9151831 PMC191659
  20. [22]Ono A Freed EO Binding of human immunodeficiency virus type 1 Gag to membrane: role of the matrix amino terminus J Virol 1999 73 4136 4144 10196310 PMC104193
  21. [23]Hermida-Matsumoto L Resh MD Localization of human immunodeficiency virus type 1 Gag and Env at the plasma membrane by confocal imaging J. Virol. 2000 74 8670 8679 10.1128/JVI.74.18.8670-8679.2000 10954568 PMC116378 · DOI
  22. [24]Ono A Freed EO Cell-type-dependent targeting of human immunodeficiency virus type 1 assembly to the plasma membrane and the multivesicular body J Virol 2004 78 1552 1563 10.1128/JVI.78.3.1552-1563.2004 14722309 PMC321403 · DOI
  23. [25]Ono A Ablan SD Lockett SJ Nagashima K Freed EO Phosphatidylinositol (4,5) bisphosphate regulates HIV-1 Gag targeting to the plasma membrane Proc. Natl. Acad. Sci. USA 2004 101 14889 14894 10.1073/pnas.0405596101 15465916 PMC522033 · DOI · Paper page
  24. [26]Chukkapalli V Hogue IB Boyko V Hu W-S Ono A Interaction between the human immunodeficiency virus type 1 Gag matrix domain and phosphatidylinositol-(4,5)-bisphosphate is essential for efficient gag membrane binding J. Virol. 2008 82 2405 2417 10.1128/JVI.01614-07 18094158 PMC2258911 · DOI · Paper page
  25. [28]Dick RA Goh SL Feigenson GW Vogt VM HIV-1 Gag protein can sense the cholesterol and acyl chain environment in model membranes Proc. Natl. Acad. Sci. USA 2012 109 18761 6 10.1073/pnas.1209408109 23010924 PMC3503231 · DOI
  26. [29]Olety B Ono A Roles played by acidic lipids in HIV-1 Gag membrane binding Virus Res. 2014 193 108 15 10.1016/j.virusres.2014.06.015 24998886 PMC4252750 · DOI
  27. [30]Mercredi, P. Y. et al. Structural and Molecular Determinants of Membrane Binding by the HIV-1 Matrix Protein. J. Mol. Biol. 10.1016/j.jmb.2016.03.005 (2016). PMC4836608 26992353 · DOI
  28. [31]Mücksch F Laketa V Müller B Schultz C Kräusslich H.-G Synchronized HIV assembly by tunable PIP2 changes reveals PIP2 requirement for stable Gag anchoring. Elife 2017 6 e25287 10.7554/eLife.25287 28574338 PMC5495570 · DOI
  29. [32]Ferrell JEJ Huestis WH Phosphoinositide metabolism and the morphology of human erythrocytes J. Cell Biol. 1984 98 1992 1998 10.1083/jcb.98.6.1992 6327723 PMC2113039 · DOI
  30. [33]Mitchell KT Ferrell JE Huestis WH Separation of phosphoinositides and other phospholipids by two-dimensional thin-layer chromatography Anal. Biochem. 1986 158 447 453 10.1016/0003-2697(86)90574-9 3028208 · DOI
  31. [34]Tran D Cellular distribution of polyphosphoinositides in rat hepatocytes Cell. Signal. 1993 5 565 581 10.1016/0898-6568(93)90052-N 8312134 · DOI
  32. [35]Stephens LR Jackson TR Hawkins PT Agonist-stimulated synthesis of phosphatidylinositol(3,4,5)-trisphosphate: A new intracellular signalling system? Biochim. Biophys. Acta - Mol. Cell Res. 1993 1179 27 75 8399352
  33. [36]Clark J Quantification of PtdInsP3 molecular species in cells and tissues by mass spectrometry Nat. Methods 2011 8 267 272 10.1038/nmeth.1564 21278744 PMC3460242 · DOI
  34. [37]Dickson EJ Hille B Understanding phosphoinositides: rare, dynamic, and essential membrane phospholipids Biochem. J. 2019 476 1 23 10.1042/BCJ20180022 30617162 PMC6342281 · DOI · Paper page
  35. [38]Welker, R., Hohenberg, H., Tessmer, U., Huckhagel, C. & Kräusslich, H. G. Biochemical and structural analysis of isolated mature cores of human immunodeficiency virus type 1. J. Virol.74, 1168–1177 (2000). PMC111451 10627527
  36. [39]Özbalci, C., Timo, S. & Brügger, B. Quantitative Analysis of Cellular Lipids by Nano-Electrospray Ionization Mass Spectrometry. Methods Mol Biol. 1033, 3–20 (2013). 23996167
  37. [40]Haag M Schmidt A Sachsenheimer T Brügger B Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) Metabolites 2012 2 57 76 10.3390/metabo2010057 24957368 PMC3901191 · DOI
  38. [41]Traynor-Kaplan A Fatty-acyl chain profiles of cellular phosphoinositides Biochim. Biophys. Acta - Mol. Cell Biol. Lipids 2017 1862 513 522 10.1016/j.bbalip.2017.02.002 28189644 PMC5392126 · DOI
  39. [42]Stephens, L., McGregor, A. & Hawkins, P. Biology of Phosphoinositides. (Oxford: Oxford University Press, 2000).
  40. [44]Charlier L Coarse-Grained Simulations of the HIV-1 Matrix Protein Anchoring: Revisiting Its Assembly on Membrane Domains Biophys. J. 2014 106 577 85 10.1016/j.bpj.2013.12.019 24507598 PMC3944987 · DOI
  41. [45]Raghupathy R Transbilayer Lipid Interactions Mediate Nanoclustering of Lipid-Anchored Proteins Cell 2015 161 581 594 10.1016/j.cell.2015.03.048 25910209 PMC4651428 · DOI
  42. [46]Skotland T Sandvig K The role of PS 18:0/18:1 in membrane function Nat. Commun. 2019 10 2752 10.1038/s41467-019-10711-1 31227693 PMC6588574 · DOI
  43. [47]Capponi S Freites JA Tobias DJ White SH Interleaflet mixing and coupling in liquid-disordered phospholipid bilayers Biochim. Biophys. Acta 2016 1858 354 362 10.1016/j.bbamem.2015.11.024 26657692 PMC4703536 · DOI
  44. [48]Huttner WB Zimmerberg J Implications of lipid microdomains for membrane curvature, budding and fission: Commentary Curr. Opin. Cell Biol. 2001 13 478 484 10.1016/S0955-0674(00)00239-8 11454455 · DOI
  45. [49]Adachi A Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone J. Virol. 1986 59 284 91 3016298 PMC253077
  46. [50]Harada S Koyanagi Y Yamamoto N Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay Science 1985 229 563 566 10.1126/science.2992081 2992081 · DOI
  47. [53]Paltauf F Hermetter A Strategies for the synthesis of glycerophospholipids Prog. Lipid Res. 1994 33 239 328 10.1016/0163-7827(94)90028-0 8022845 · DOI
  48. [54]Kielkowska A A new approach to measuring phosphoinositides in cells by mass spectrometry Adv. Biol. Regul. 2014 54 131 141 10.1016/j.jbior.2013.09.001 24120934 · DOI

Cited by

Sorted by citations

Authors
Mücksch, F., Citir, M., Lüchtenborg, C., Glass, B., Traynor-Kaplan, A., Schultz, C., Brügger, B., & Kräusslich, H.-G.
Affiliations
  • European Molecular Biology Laboratory (DE)
  • Heidelberg University (DE)
  • University Hospital Heidelberg (DE)
  • Oregon Health & Science University (US)
Corresponding author
Kräusslich, Hans-Georg
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2019-11-27
Volume
9
Issue
1
Pages
17661-17661
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-019-53939-z
PMC
PMC6881329
OpenAlex
W2990786400
Cited by
73
References
6
Citations by year
2026: 7 · 2025: 12 · 2024: 12 · 2023: 8 · 2022: 9 · 2021: 14 · 2020: 11
FWCI
4.76
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Deutsches Zentrum für Infektionsforschung (Project 7.5 TTU HIV)
  • Deutsche Forschungsgemeinschaft (TRR186, 240245660, SFB1129, 278001972, 316659730, 112927078, TRR 83)
Topics
  • Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
Keywords
HIV-1, phosphoinositides, phosphatidylinositol 4,5-bisphosphate, phosphatidylinositol 3,4,5-trisphosphate, lipid rafts, Gag protein, viral budding, plasma membrane lipid composition, lipidomics
MeSH terms
Cell Membrane, Humans, Lipids, Phosphatidylinositols, HIV-1, Virus Assembly, Phosphatidylinositol 4,5-Diphosphate, Membrane Microdomains, gag Gene Products, Human Immunodeficiency Virus, Virus Release

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