Immunoelectron Microscopic Evidence for Tetherin/BST2 as the Physical Bridge between HIV-1 Virions and the Plasma Membrane

English version České info

Tetherin/BST2 was identified in 2008 as the cellular factor responsible for restricting HIV-1 replication at a very late stage in the lifecycle. Tetherin acts to retain virion particles on the plasma membrane after budding has been completed. Infected cells that express large amounts of tetherin display large strings of HIV virions that remain attached to the plasma membrane. Vpu is an HIV-1 accessory protein that specifically counteracts the restriction to virus release contributed by tetherin. Tetherin is an unusual Type II transmembrane protein that contains a GPI anchor at its C-terminus and is found in lipid rafts. The leading model for the mechanism of action of tetherin is that it functions as a direct physical tether bridging virions and the plasma membrane. However, evidence that tetherin functions as a physical tether has thus far been indirect. Here we demonstrate by biochemical and immunoelectron microscopic methods that endogenous tetherin is present on the viral particle and forms a bridge between virion particles and the plasma membrane. Endogenous tetherin was found on HIV particles that were released by partial proteolytic digestion. Immunoelectron microscopy performed on HIV-infected T cells demonstrated that tetherin forms an apparent physical link between virions and connects patches of virions to the plasma membrane. Linear filamentous strands that were highly enriched in tetherin bridged the space between some virions. We conclude that tetherin is the physical tether linking HIV-1 virions and the plasma membrane. The presence of filaments with which multiple molecules of tetherin interact in connecting virion particles is strongly suggested by the morphologic evidence.

Vyšlo v časopise: Immunoelectron Microscopic Evidence for Tetherin/BST2 as the Physical Bridge between HIV-1 Virions and the Plasma Membrane. PLoS Pathog 6(2): e32767. doi:10.1371/journal.ppat.1000749
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000749

Souhrn

Zdroje

1. McDonaldB

Martin-SerranoJ

2009 No strings attached: the ESCRT machinery in viral budding and cytokinesis. J Cell Sci 122 2167 2177

2. StuchellMD

GarrusJE

MullerB

StrayKM

GhaffarianS

2004 The human endosomal sorting complex required for transport (ESCRT-I) and its role in HIV-1 budding. J Biol Chem 279 36059 36071

3. GoffSP

2004 Retrovirus restriction factors. Mol Cell 16 849 859

4. YuX

YuY

LiuB

LuoK

KongW

2003 Induction of APOBEC3G ubiquitination and degradation by an HIV-1 Vif-Cul5-SCF complex. Science 302 1056 1060

5. BieniaszPD

2004 Intrinsic immunity: a front-line defense against viral attack. Nat Immunol 5 1109 1115

6. SebastianS

LubanJ

2007 The Retroviral Restriction Factor TRIM5alpha. Curr Infect Dis Rep 9 167 173

7. TowersGJ

2007 The control of viral infection by tripartite motif proteins and cyclophilin A. Retrovirology 4 40

8. NeilSJ

ZangT

BieniaszPD

2008 Tetherin inhibits retrovirus release and is antagonized by HIV-1 Vpu. Nature 451 425 430

9. Van DammeN

GoffD

KatsuraC

JorgensonRL

MitchellR

2008 The interferon-induced protein BST-2 restricts HIV-1 release and is downregulated from the cell surface by the viral Vpu protein. Cell Host Microbe 3 245 252

10. StrebelK

KlimkaitT

MartinMA

1988 A novel gene of HIV-1, vpu, and its 16-kilodalton product. Science 241 1221 1223

11. CohenEA

TerwilligerEF

SodroskiJG

HaseltineWA

1988 Identification of a protein encoded by the vpu gene of HIV-1. Nature 334 532 534

12. WilleyRL

MaldarelliF

MartinMA

StrebelK

1992 Human immunodeficiency virus type 1 Vpu protein regulates the formation of intracellular gp160-CD4 complexes. J Virol 66 226 234

13. MargottinF

BourSP

DurandH

SeligL

BenichouS

1998 A novel human WD protein, h-beta TrCp, that interacts with HIV-1 Vpu connects CD4 to the ER degradation pathway through an F-box motif. Mol Cell 1 565 574

14. WilleyRL

MaldarelliF

MartinMA

StrebelK

1992 Human immunodeficiency virus type 1 Vpu protein induces rapid degradation of CD4. J Virol 66 7193 7200

15. KlimkaitT

StrebelK

HogganMD

MartinMA

OrensteinJM

1990 The human immunodeficiency virus type 1-specific protein vpu is required for efficient virus maturation and release. J Virol 64 621 629

16. VarthakaviV

SmithRM

BourSP

StrebelK

SpearmanP

2003 Viral protein U counteracts a human host cell restriction that inhibits HIV-1 particle production. Proc Natl Acad Sci U S A 100 15154 15159

17. SchubertU

StrebelK

1994 Differential activities of the human immunodeficiency virus type 1-encoded Vpu protein are regulated by phosphorylation and occur in different cellular compartments. J Virol 68 2260 2271

18. SchubertU

BourS

Ferrer-MontielAV

MontalM

MaldarellF

1996 The two biological activities of human immunodeficiency virus type 1 Vpu protein involve two separable structural domains. J Virol 70 809 819

19. GottlingerHG

DorfmanT

CohenEA

HaseltineWA

1993 Vpu protein of human immunodeficiency virus type 1 enhances the release of capsids produced by gag gene constructs of widely divergent retroviruses. Proc Natl Acad Sci U S A 90 7381 7385

20. GeraghtyRJ

TalbotKJ

CallahanM

HarperW

PanganibanAT

1994 Cell type-dependence for Vpu function. J Med Primatol 23 146 150

21. SakaiH

TokunagaK

KawamuraM

AdachiA

1995 Function of human immunodeficiency virus type 1 Vpu protein in various cell types. J Gen Virol 76(Pt 11) 2717 2722

22. KupzigS

KorolchukV

RollasonR

SugdenA

WildeA

2003 Bst-2/HM1.24 is a raft-associated apical membrane protein with an unusual topology. Traffic 4 694 709

23. DubeM

RoyBB

Guiot-GuillainP

MercierJ

BinetteJ

2009 Suppression of Tetherin-restricting activity upon human immunodeficiency virus type 1 particle release correlates with localization of Vpu in the trans-Golgi network. J Virol 83 4574 4590

24. MitchellRS

KatsuraC

SkaskoMA

FitzpatrickK

LauD

2009 Vpu antagonizes BST-2-mediated restriction of HIV-1 release via beta-TrCP and endo-lysosomal trafficking. PLoS Pathog 5 e1000450 doi:10.1371/journal.ppat.1000450

25. GoffinetC

AllespachI

HomannS

TervoHM

HabermannA

2009 HIV-1 antagonism of CD317 is species specific and involves Vpu-mediated proteasomal degradation of the restriction factor. Cell Host Microbe 5 285 297

26. MiyagiE

AndrewAJ

KaoS

StrebelK

2009 Vpu enhances HIV-1 virus release in the absence of Bst-2 cell surface down-modulation and intracellular depletion. Proc Natl Acad Sci U S A 106 2868 2873

27. KaletskyRL

FrancicaJR

Agrawal-GamseC

BatesP

2009 Tetherin-mediated restriction of filovirus budding is antagonized by the Ebola glycoprotein. Proc Natl Acad Sci U S A 106 2886 2891

28. SakumaT

NodaT

UrataS

KawaokaY

YasudaJ

2009 Inhibition of Lassa and Marburg virus production by tetherin. J Virol 83 2382 2385

29. JouvenetN

NeilSJ

ZhadinaM

ZangT

KratovacZ

2009 Broad-spectrum inhibition of retroviral and filoviral particle release by tetherin. J Virol 83 1837 1844

30. ZhangF

WilsonSJ

LandfordWC

VirgenB

GregoryD

2009 Nef Proteins from Simian Immunodeficiency Viruses Are Tetherin Antagonists. Cell Host Microbe

31. BourS

StrebelK

1996 The human immunodeficiency virus (HIV) type 2 envelope protein is a functional complement to HIV type 1 Vpu that enhances particle release of heterologous retroviruses. J Virol 70 8285 8300

32. DouglasJL

ViswanathanK

McCarrollMN

GustinJK

FruhK

2009 Vpu Directs the Degradation of the HIV Restriction Factor BST-2/tetherin via a {beta}TrCP-dependent Mechanism. J Virol

33. Perez-CaballeroD

ZangT

EbrahimiA

McNattMW

GregoryDA

2009 Tetherin inhibits HIV-1 release by directly tethering virions to cells. Cell 139 499 511

34. RollasonR

KorolchukV

HamiltonC

JepsonM

BantingG

2009 A CD317/tetherin-RICH2 complex plays a critical role in the organization of the subapical actin cytoskeleton in polarized epithelial cells. J Cell Biol 184 721 736

35. YeeJK

FriedmannT

BurnsJC

1994 Generation of high-titer pseudotyped retroviral vectors with very broad host range. Methods Cell Biol 43 Pt A 99 112

36. MorgensternJP

LandH

1990 Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res 18 3587 3596

37. SandefurS

SmithRM

VarthakaviV

SpearmanP

2000 Mapping and characterization of the N-terminal I domain of human immunodeficiency virus type 1 Pr55(Gag). J Virol 74 7238 7249