Effects of Microtubule Modulators on HIV-1 Infection of Transformed and Resting CD4 T Cells

Yoder, Alyson; Guo, Jia; Yu, Dongyang; Cui, Zongqiang; Zhang, Xian-En; Wu, Yuntao

doi:10.1128/jvi.02462-10

Cited by 35 publications

(33 citation statements)

References 49 publications

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“…Sabo et al recently reported that stable microtubules facilitate early events in infection (13). However, another study observed no effect on infectivity when microtubules were disrupted using Noc (26). The data presented here are consistent with all three studies.…”

Section: Discussionsupporting

confidence: 82%

“…In the presence of CsA, infection was only modestly affected by Noc treatment (Fig. 1C), in agreement with a previous report that transient Noc treatment does not inhibit HIV-1 infection (26). Transient Noc treatment did not disrupt the ability of TRIM-Cyp to restrict HIV-1 infection (Fig.…”

Section: Microtubule Disruption Delays the Uncoating Kinetics Of Hiv-1supporting

confidence: 80%

“…Similar results were observed upon Noc treatment for 2 h (data not shown). This demonstrates that Noc treatment does not directly influence RT in these infections, since any differences observed occurred well after the removal of Noc, a finding supported by similar findings for CD4 ϩ T cells by Yoder et al (26). The fate-of-capsid assay confirms microtubule-dependent uncoating of HIV-1.…”

Section: Microtubule Disruption Delays the Uncoating Kinetics Of Hiv-1supporting

confidence: 75%

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HIV-1 Uncoating Is Facilitated by Dynein and Kinesin 1

Lukic

Dharan

Fricke

et al. 2014

J Virol

114

150

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Following entry into the target cell, human immunodeficiency virus type 1 (HIV-1) must reverse transcribe its RNA genome to DNA and traffic to the nuclear envelope, where the viral genome is translocated into the nucleus for subsequent integration into the host cell chromosome. During this time, the viral core, which houses the genome, undergoes a poorly understood process of disassembly, known as uncoating. Collectively, many studies suggest that uncoating is tightly regulated to allow nuclear import of the genome while minimizing the exposure of the newly synthesized DNA to cytosolic DNA sensors. However, whether host cellular proteins facilitate this process remains poorly understood. Here we report that intact microtubules facilitate HIV-1 uncoating in target cells. Disruption of microtubules with nocodazole substantially delays HIV-1 uncoating, as revealed with three different assay systems. This defect in uncoating did not correlate with defective reverse transcription at early times postinfection, demonstrating that microtubule-facilitated uncoating is distinct from the previously reported role of viral reverse transcription in the uncoating process. We also find that pharmacological or small interfering RNA (siRNA)-mediated inhibition of cytoplasmic dynein or the kinesin 1 heavy chain KIF5B delays uncoating, providing detailed insight into how microtubules facilitate the uncoating process. These studies reveal a previously unappreciated role for microtubules and microtubule motor function in HIV-1 uncoating, establishing a functional link between viral trafficking and uncoating. Targeted disruption of the capsid motor interaction may reveal novel mechanisms of inhibition of viral infection or provide opportunities to activate cytoplasmic antiviral responses directed against capsid or viral DNA. IMPORTANCEDuring HIV-1 infection, fusion of viral and target cell membranes dispenses the viral ribonucleoprotein complex into the cytoplasm of target cells. During this time, the virus must reverse transcribe its RNA genome, traffic from the location of fusion to the nuclear membrane, and undergo the process of uncoating, whereby the viral capsid core disassembles to allow the subsequent nuclear import of the viral genome. Numerous cellular restriction factors target the viral capsid, suggesting that perturbation of the uncoating process represents an excellent antiviral target. However, this uncoating process, and the cellular factors that facilitate uncoating, remains poorly understood. The main observation of this study is that normal uncoating requires intact microtubules and is facilitated by dynein and kinesin motors. Targeting these factors may either directly inhibit infection or delay it enough to trigger mediators of intrinsic immunity that recognize cytoplasmic capsid or DNA and subsequently induce an antiviral state in these cells.

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Section: Discussionsupporting

confidence: 82%

Section: Microtubule Disruption Delays the Uncoating Kinetics Of Hiv-1supporting

confidence: 80%

Section: Microtubule Disruption Delays the Uncoating Kinetics Of Hiv-1supporting

confidence: 75%

See 1 more Smart Citation

HIV-1 Uncoating Is Facilitated by Dynein and Kinesin 1

Lukic

Dharan

Fricke

et al. 2014

J Virol

114

150

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“…SUN2 overexpression inhibited single-round infection equally in the presence of control treatment (dimethyl sulfoxide [DMSO]) or nocodazole (3.3ϫ and 3.5ϫ blocks to single-round infection, respectively), which suggests that intact microtubules are not required for the SUN2-imposed block to infection. While the effect of nocodazole on HIV infection that we observed is greater than that often reported (66,69,70), a similar 2-to 3-fold effect also has been described (69,71), including in CHME cells (62), which could reflect cell typedependent effects or experimental variation (e.g., whether nocodazole was removed or maintained during infection).…”

Section: Fig 3 Differential Sensitivity Of Hiv Strains To Sun2 Contrcontrasting

confidence: 41%

SUN2 Overexpression Deforms Nuclear Shape and Inhibits HIV

Donahue

Amraoui

Nunzio

et al. 2016

J Virol

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In a previous screen of putative interferon-stimulated genes, SUN2 was shown to inhibit HIV-1 infection in an uncharacterized manner. SUN2 is an inner nuclear membrane protein belonging to the linker of nucleoskeleton and cytoskeleton complex. We have analyzed here the role of SUN2 in HIV infection. We report that in contrast to what was initially thought, SUN2 is not induced by type I interferon, and that SUN2 silencing does not modulate HIV infection. However, SUN2 overexpression in cell lines and in primary monocyte-derived dendritic cells inhibits the replication of HIV but not murine leukemia virus or chikungunya virus. We identified HIV-1 and HIV-2 strains that are unaffected by SUN2, suggesting that the effect is specific to particular viral components or cofactors. Intriguingly, SUN2 overexpression induces a multilobular flower-like nuclear shape that does not impact cell viability and is similar to that of cells isolated from patients with HTLV-I-associated adult T-cell leukemia or with progeria. Nuclear shape changes and HIV inhibition both mapped to the nucleoplasmic domain of SUN2 that interacts with the nuclear lamina. This block to HIV replication occurs between reverse transcription and nuclear entry, and passaging experiments selected for a single-amino-acid change in capsid (CA) that leads to resistance to overexpressed SUN2. Furthermore, using chemical inhibition or silencing of cyclophilin A (CypA), as well as CA mutant viruses, we implicated CypA in the SUN2-imposed block to HIV infection. Our results demonstrate that SUN2 overexpression perturbs both nuclear shape and early events of HIV infection. IMPORTANCECells encode proteins that interfere with viral replication, a number of which have been identified in overexpression screens. SUN2 is a nuclear membrane protein that was shown to inhibit HIV infection in such a screen, but how it blocked HIV infection was not known. We show that SUN2 overexpression blocks the infection of certain strains of HIV before nuclear entry. Mutation of the viral capsid protein yielded SUN2-resistant HIV. Additionally, the inhibition of HIV infection by SUN2 involves cyclophilin A, a protein that binds the HIV capsid and directs subsequent steps of infection. We also found that SUN2 overexpression substantially changes the shape of the cell's nucleus, resulting in many flower-like nuclei. Both HIV inhibition and deformation of nuclear shape required the domain of SUN2 that interacts with the nuclear lamina. Our results demonstrate that SUN2 interferes with HIV infection and highlight novel links between nuclear shape and viral infection.

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“…Microtubules and Myosin II Contribute to FFWO-We and others have shown that HIV-1 fused efficiently with target cells pretreated with microtubule depolymerizing drugs (13,51). To assess the role of microtubules in FFWO, DSP-1 and DSP-2 cells were pretreated with nocodazole (to depolymerize microtubules) or with paclitaxel (to stabilize microtubules), and the ability of HIV-1 pseudoparticles to mediate cell-cell fusion was evaluated.…”

Section: Measurements Of Virus-cell Fusion and Virus-mediated Cellcelmentioning

confidence: 99%