Critical role for the kinesin KIF3A in the HIV life cycle in primary human macrophages

Gaudin, Raphaël; Alencar, Bruna Cunha de; Jouve, Mabel; Berrè, Stefano; Bouder, Emmanuel Le; Schindler, Michael; Varthaman, Aditi; Gobert, François-Xavier; Benaroch, Philippe

doi:10.1083/jcb.201201144

Cited by 43 publications

(47 citation statements)

References 43 publications

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“…In this present study, we found that eATP represents a microenvironmental stimulus capable of inducing rapid discharge of VCCassociated virions from infected macrophages. Given that at least one-third of VCC have been reported to be connected to the plasma membrane (11,21), the gross morphological changes associated with eATP stimulation, resulting in cell contraction and membrane blebbing, could lead to an increased tension of the plasma membrane. VCC may then open up, bringing important amounts of membranes to the plasma membrane and thereby reducing the tension along with the release of the VCC content in the extracellular milieu.…”

Section: Discussionmentioning

confidence: 99%

“…More recently, VCC are believed to represent intracellular sequestration of plasma membrane areas rich in a subset of tetraspanins (12). Likely because of their origin, VCC can be transiently (11) connected to the extracellular medium through microchannels or conduits (21,22). Of interest, VCC-like compartments accessible by the extracellular medium preexist in uninfected macrophages and express similar markers, including CD9, CD81, CD63 (23), and, identified more recently, CD36 (24).…”

Section: Hiv Type 1 (Hiv-1) Infects Cd4mentioning

confidence: 99%

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Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages

Graziano

Desdouits

Garzetti

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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HIV type 1 (HIV-1) infects CD4+ T lymphocytes and tissue macrophages. Infected macrophages differ from T cells in terms of decreased to absent cytopathicity and for active accumulation of new progeny HIV-1 virions in virus-containing compartments (VCC). For these reasons, infected macrophages are believed to act as “Trojan horses” carrying infectious particles to be released on cell necrosis or functional stimulation. Here we explored the hypothesis that extracellular ATP (eATP) could represent a microenvironmental signal potentially affecting virion release from VCC of infected macrophages. Indeed, eATP triggered the rapid release of infectious HIV-1 from primary human monocyte-derived macrophages (MDM) acutely infected with the CCR5-dependent HIV-1 strain. A similar phenomenon was observed in chronically infected promonocytic U1 cells differentiated to macrophage-like cells (D-U1) by costimulation with phorbol esters and urokinase-type plasminogen activator. Worthy of note, eATP did not cause necrotic, apoptotic, or pyroptotic cell death, and its effect on HIV-1 release was suppressed by Imipramine (an antidepressant agent known to inhibit microvesicle formation by interfering with membrane-associated acid sphingomyelinase). Virion release was not triggered by oxidized ATP, whereas the effect of eATP was inhibited by a specific inhibitor of the P2X7 receptor (P2X7R). Thus, eATP triggered the discharge of virions actively accumulating in VCC of infected macrophages via interaction with the P2X7R in the absence of significant cytopathicity. These findings suggest that the microvesicle pathway and P2X7R could represent exploitable targets for interfering with the VCC-associated reservoir of infectious HIV-1 virions in tissue macrophages.

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Section: Discussionmentioning

confidence: 99%

Section: Hiv Type 1 (Hiv-1) Infects Cd4mentioning

confidence: 99%

Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages

Graziano

Desdouits

Garzetti

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…These compartments are termed intracellular PM-connected compartments (IPMCs) (24). Previous studies have also suggested the involvement of host cellular components, such as kinesins, in formation and/or dynamics of VCCs or IPMCs (27,28).…”

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confidence: 99%

Molecular Determinants Directing HIV-1 Gag Assembly to Virus-Containing Compartments in Primary Macrophages

Inlora

Chukkapalli

Bedi

et al. 2016

J Virol

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The subcellular sites of HIV-1 assembly, determined by the localization of the structural protein Gag, vary in a cell-type-dependent manner. In T cells and transformed cell lines used as model systems, HIV-1 assembles at the plasma membrane (PM). The binding and localization of HIV-1 Gag to the PM are mediated by the interaction between the matrix (MA) domain, specifically the highly basic region, and a PM-specific acidic phospholipid, phosphatidylinositol-4,5-bisphosphate [PI(4,5)P 2 ]. In primary macrophages, prominent accumulation of assembling or assembled particles is found in the virus-containing compartments (VCCs), which largely consist of convoluted invaginations of the PM. To elucidate the molecular mechanism of HIV-1 Gag targeting to the VCCs, we examined the impact of overexpression of polyphosphoinositide 5-phosphatase IV (5ptaseIV), which depletes cellular PI(4,5)P 2 , in primary macrophages. We found that the VCC localization and virus release of HIV-1 are severely impaired upon 5ptaseIV overexpression, suggesting an important role for the MA-PI(4,5)P 2 interaction in HIV-1 assembly in primary macrophages. However, our analysis of HIV-1 Gag derivatives with MA changes showed that this interaction contributes to Gag membrane binding but is dispensable for specific targeting of Gag to the VCCs per se. We further determined that deletion of the NC domain abolishes VCC-specific localization of HIV-1 Gag. Notably, HIV-1 Gag localized efficiently to the VCCs when the NC domain was replaced with a leucine zipper dimerization motif that promotes Gag multimerization. Altogether, our data revealed that targeting of HIV-1 Gag to the VCCs requires NC-dependent multimerization. IMPORTANCEIn T cells and model cell lines, HIV-1 Gag localizes to the PM in a manner dependent on the MA-PI(4,5)P 2 interaction. On the other hand, in primary macrophages, HIV-1 Gag localizes to convoluted intracellular membrane structures termed virus-containing compartments (VCCs). Although these compartments have been known for decades, and despite the implication of viruses in VCCs being involved in virus reservoir maintenance and spread, the viral determinant(s) that promotes Gag targeting to VCCs is unknown. In this study, we found that the MA-PI(4,5)P 2 interaction facilitates efficient Gag membrane binding in macrophages but is not essential for Gag targeting to VCCs. Rather, our results revealed that NC-dependent multimerization promotes VCC targeting. Our findings highlight the differential roles played by MA and NC in HIV-1 Gag membrane binding and targeting and suggest a multimerization-dependent mechanism for Gag trafficking in primary macrophages similar to that for Gag localization to uropods in polarized T cells.

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“…With the advent of superresolution microscopy, it became possible to examine viral processes unhindered by the diffraction limit. The initial studies using PALM (photoactivated localization microscopy) documented Gag puncta at the plasma membrane (44), while more-recent analyses focused on HIV-1 entry and trafficking (45,46) and HIV-1 Gag assembly (47)(48)(49)(50)(51) and revealed the localization of the restriction factor tetherin relative to Gag and Env (52). Finally, a very recent study, which provided subdiffraction mapping of Gag and Env at the plasma membrane (53), along with elegant correlative electron/fluorescence microscopy (54,55), addresses the specificity of Env recruitment and its incorporation into budding particles.…”

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confidence: 99%

Clustering and Mobility of HIV-1 Env at Viral Assembly Sites Predict Its Propensity To Induce Cell-Cell Fusion

Roy

Chan

Lambelé

et al. 2013

J Virol

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HIV-1 Env mediates virus attachment to and fusion with target cell membranes, and yet, while Env is still situated at the plasma membrane of the producer cell and before its incorporation into newly formed particles, Env already interacts with the viral receptor CD4 on target cells, thus enabling the formation of transient cell contacts that facilitate the transmission of viral particles. During this first encounter with the receptor, Env must not induce membrane fusion, as this would prevent the producer cell and the target cell from separating upon virus transmission, but how Env's fusion activity is controlled remains unclear. To gain a better understanding of the Env regulation that precedes viral transmission, we examined the nanoscale organization of Env at the surface of producer cells. Utilizing superresolution microscopy (stochastic optical reconstruction microscopy [STORM]) and fluorescence recovery after photobleaching (FRAP), we quantitatively assessed the clustering and dynamics of Env upon its arrival at the plasma membrane. We found that Gag assembly induced the aggregation of small Env clusters into larger domains and that these domains were completely immobile. Truncation of the cytoplasmic tail (CT) of Env abrogated Gag's ability to induce Env clustering and restored Env mobility at assembly sites, both of which correlated with increased Env-induced fusion of infected and uninfected cells. Hence, while Env trapping by Gag secures Env incorporation into viral particles, Env clustering and its sequestration at assembly sites likely also leads to the repression of its fusion function, and thus, by preventing the formation of syncytia, Gag helps to secure efficient transfer of viral particles to target cells.

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Critical role for the kinesin KIF3A in the HIV life cycle in primary human macrophages

Abstract: KIF3A is important for the intracellular transport of HIV-containing compartments and HIV release from infected macrophages.

Cited by 43 publications

References 43 publications

Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages

Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages

Molecular Determinants Directing HIV-1 Gag Assembly to Virus-Containing Compartments in Primary Macrophages

Clustering and Mobility of HIV-1 Env at Viral Assembly Sites Predict Its Propensity To Induce Cell-Cell Fusion

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