C-terminal Hydrophobic Region in Human Bone Marrow Stromal Cell Antigen 2 (BST-2)/Tetherin Protein Functions as Second Transmembrane Motif

Andrew, Amy J.; Kao, Sandra; Strebel, Klaus

doi:10.1074/jbc.m111.287011

Cited by 35 publications

(44 citation statements)

References 52 publications

(63 reference statements)

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“…Another line of evidence in support of this hypothesis is the presence of tetherin in Triton X-insoluble membrane fractions (16,18). Even though a study of tetherin chimeras in which the GPI anchor is replaced with a heterologous transmembrane domain showed that lipid raft association measured by detergent resistance is not sufficient (84), it was possible that association of tetherin with lipid rafts may play an important role. In the present study, however, using an antibody copatching assay, we found that tetherin itself does not appear to associate strongly with multiple lipid raft markers (CD55, CD59, NFP-GPI, and NFP-HATM) at the plasma membrane, all of which were previously shown to copatch strongly with WT Gag (26).…”

Section: Discussionmentioning

confidence: 80%

Roles Played by Capsid-Dependent Induction of Membrane Curvature and Gag-ESCRT Interactions in Tetherin Recruitment to HIV-1 Assembly Sites

Grover

Llewellyn

Soheilian

et al. 2013

J Virol

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Tetherin/BST-2 (here called tetherin) is an antiviral protein that restricts release of diverse enveloped viruses from infected cells through physically tethering virus envelope and host plasma membrane. For HIV-1, specific recruitment of tetherin to assembly sites has been observed as its colocalization with the viral structural protein Gag or its accumulation in virus particles. Because of its broad range of targets, we hypothesized that tetherin is recruited through conserved features shared among various enveloped viruses, such as lipid raft association, membrane curvature, or ESCRT dependence. We observed that reduction of cellular cholesterol does not block tetherin anti-HIV-1 function, excluding an essential role for lipid rafts. In contrast, mutations in the capsid domain of Gag, which inhibit induction of membrane curvature, prevented tetherin-Gag colocalization detectable by confocal microscopy. Disruption of Gag-ESCRT interactions also inhibited tetherin-Gag colocalization when disruption was accomplished via amino acid substitutions in late domain motifs, expression of a dominant-negative Tsg101 derivative, or small interfering RNA (siRNA)-mediated depletion of Tsg101 or Alix. However, further analyses of these conditions by quantitative superresolution localization microscopy revealed that Gag-tetherin coclustering is significantly reduced but persists at intermediate levels. Notably, this residual tetherin recruitment was still sufficient for the full restriction of HIV-1 release. Unlike the late domain mutants, the capsid mutants defective in inducing membrane curvature showed little or no coclustering with tetherin in superresolution analyses. These results support a model in which both Gag-induced membrane curvature and Gag-ESCRT interactions promote tetherin recruitment, but the recruitment level achieved by the former is sufficient for full restriction.

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Section: Discussionmentioning

confidence: 80%

Roles Played by Capsid-Dependent Induction of Membrane Curvature and Gag-ESCRT Interactions in Tetherin Recruitment to HIV-1 Assembly Sites

Grover

Llewellyn

Soheilian

et al. 2013

J Virol

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“…Plasmid pcDNA-BST-2 is a vector for the expression of human BST-2 under control of the cytomegalovirus immediate-early promoter (29). HA-tagged BST-2 encoding a triple HA epitope tag in the ectodomain following BST-2 residue 148 (BST-2 I ) was constructed using PCR-based methodologies as described (24). BST-2 I is referred to here as BST-2 wt and was used for construction of additional mutants.…”

Section: Methodsmentioning

confidence: 99%

“…For consistency with immune recognition, we utilized a BST-2 construct containing a triple HA tag downstream of residue 148 near the C terminus of the ectodomain as described previously (24). We refer to this construct as BST-2 wt for the remainder of the study.…”

Section: Replacing the C-terminal Coiled-coil Domain With A Noncoiledmentioning

confidence: 99%

“…The protein is predicted to have an N-terminal transmembrane (TM) 2 region and a C-terminal glycosylphosphatidylinositol anchor (23). However, we recently found evidence to suggest that the C-terminal membrane anchor could be a second transmembrane domain rather than a glycosylphosphatidylinositol anchor (24). The two TM domains are separated by ϳ120 residues that constitute the ectodomain of the protein, which is predicted to form a coiled-coil structure (25)(26)(27)(28).…”

mentioning

confidence: 99%

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The Size and Conservation of a Coiled-coil Structure in the Ectodomain of Human BST-2/Tetherin Is Dispensable for Inhibition of HIV-1 Virion Release

Andrew

Berndsen

Kao

et al. 2012

Journal of Biological Chemistry

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Background: BST-2 inhibits virus release by tethering virions to the cell surface. Results: The length of the BST-2 ectodomain can be increased or reduced without loss of function. Conclusion:The positioning of ectodomain deletions rather than their size determines the impact on BST-2 function. Significance: Understanding the importance of structural elements in BST-2 is critical for developing antiviral strategies.

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“…The BST-2 C terminus is made up of a stretch of hydrophobic residues that, in rat BST-2, has been determined to function as a glycosylphosphatidylinositol (GPI) anchor signal (4). Recent experimental evidence suggests that in human BST-2 (huBST-2), this C-terminal hydrophobic domain may constitute a second TM domain rather than a GPI anchor signal (9). As far as the BST-2 ectodomain is concerned, it contains three cysteine residues important for the formation of covalent cysteine-linked dimers, as well as two N-linked carbohydrate side chains whose functional importance is currently unclear (1, 2, 4, 10, 11).…”

mentioning

confidence: 99%

Functional Antagonism of Rhesus Macaque and Chimpanzee BST-2 by HIV-1 Vpu Is Mediated by Cytoplasmic Domain Interactions

Yoshida

Koyanagi

Strebel

2013

J Virol

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bHuman immunodeficiency virus type 1 (HIV-1) Vpu enhances the release of viral particles from infected cells by interfering with the function of BST-2/tetherin, a cellular protein inhibiting virus release. The Vpu protein encoded by NL4-3, a widely used HIV-1 laboratory strain, antagonizes human BST-2 but not monkey or murine BST-2, leading to the conclusion that BST-2 antagonism by Vpu is species specific. In contrast, we recently identified several primary Vpu isolates, such as Vpu of HIV-1 DH12 , capable of antagonizing both human and rhesus BST-2. Here we report that while Vpu interacts with human BST-2 primarily through their respective transmembrane domains, antagonism of rhesus BST-2 by Vpu involved an interaction of their cytoplasmic domains. Importantly, a Vpu mutant carrying two mutations in its transmembrane domain (A 14 L and W 22 A), rendering it incompetent for interaction with human BST-2, was able to interact with human BST-2 carrying the rhesus BST-2 cytoplasmic domain and partially neutralized the ability of this BST-2 variant to inhibit viral release. Bimolecular fluorescence complementation analysis to detect Vpu-BST-2 interactions suggested that the physical interaction of Vpu with rhesus or chimpanzee BST-2 involves a 5-residue motif in the cytoplasmic domain of BST-2 previously identified as important for the antagonism of monkey and great ape BST-2 by simian immunodeficiency virus (SIV) Nef. Thus, our study identifies a novel mechanism of antagonism of monkey and great ape BST-2 by Vpu that targets the same motif in BST-2 used by SIV Nef and might explain the expanded host range observed for Vpu isolates in our previous study.

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C-terminal Hydrophobic Region in Human Bone Marrow Stromal Cell Antigen 2 (BST-2)/Tetherin Protein Functions as Second Transmembrane Motif

Cited by 35 publications

References 52 publications

Roles Played by Capsid-Dependent Induction of Membrane Curvature and Gag-ESCRT Interactions in Tetherin Recruitment to HIV-1 Assembly Sites

Roles Played by Capsid-Dependent Induction of Membrane Curvature and Gag-ESCRT Interactions in Tetherin Recruitment to HIV-1 Assembly Sites

The Size and Conservation of a Coiled-coil Structure in the Ectodomain of Human BST-2/Tetherin Is Dispensable for Inhibition of HIV-1 Virion Release

Functional Antagonism of Rhesus Macaque and Chimpanzee BST-2 by HIV-1 Vpu Is Mediated by Cytoplasmic Domain Interactions

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