Membrane metalloprotease TRABD2A restricts HIV-1 progeny production in resting CD4+ T cells by degrading viral Gag polyprotein

Liang, Guoxin; Zhao, Li; Qiao, Ying; Geng, Wenqing; Zhang, Xiaowei; Liu, Mei; Dong, Jinxiu; Ding, Haibo; Sun, Hong; Shang, Hong

doi:10.1038/s41590-019-0385-2

Cited by 19 publications

(21 citation statements)

References 41 publications

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“…In fact, we observed that Nef-LLAA is unable to target HDAC6 and to neutralize HDAC6-mediated Pr55Gag degradation and inhibition of viral production and infection. This anti-HIV-1 activity of HDAC6 against Pr55Gag could be very important, and points to a similar action exerted by the cell-membrane metalloprotease TRAB domain-containing protein 2A (TRABD2A), which acts on resting CD4+ T-cells (Liang et al, 2019). Of note, HDAC6 also controls HIV-1 infectiveness by targeting Vif, as we observed and reported here (Valera et al, 2015).…”

Section: Discussionmentioning

confidence: 80%

HIV-1 Nef Targets HDAC6 to Assure Viral Production and Virus Infection

et al. 2019

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HIV Nef is a central auxiliary protein in HIV infection and pathogenesis. Our results indicate that HDAC6 promotes the aggresome/autophagic degradation of the viral polyprotein Pr55Gag to inhibit HIV-1 production. Nef counteracts this antiviral activity of HDAC6 by inducing its degradation and subsequently stabilizing Pr55Gag and Vif viral proteins. Nef appears to neutralize HDAC6 by an acidic/endosomal-lysosomal processing and does not need the downregulation function, since data obtained with the non-associated cell-surface Nef-G2A mutant – the cytoplasmic location of HDAC6 – together with studies with chemical inhibitors and other Nef mutants, point to this direction. Hence, the polyproline rich region P72xxP75 (69–77 aa) and the di-Leucin motif in the Nef-ExxxLL160-165 sequence of Nef, appear to be responsible for HDAC6 clearance and, therefore, required for this novel Nef proviral function. Nef and Nef-G2A co-immunoprecipitate with HDAC6, whereas the Nef-PPAA mutant showed a reduced interaction with the anti-HIV-1 enzyme. Thus, the P72xxP75 motif appears to be responsible, directly or indirectly, for the interaction of Nef with HDAC6. Remarkably, by neutralizing HDAC6, Nef assures Pr55Gag location and aggregation at plasma membrane, as observed by TIRFM, promotes viral egress, and enhances the infectivity of viral particles. Consequently, our results suggest that HDAC6 acts as an anti-HIV-1 restriction factor, limiting viral production and infection by targeting Pr55Gag and Vif. This function is counteracted by functional HIV-1 Nef, in order to assure viral production and infection capacities. The interplay between HIV-1 Nef and cellular HDAC6 may determine viral infection and pathogenesis, representing both molecules as key targets to battling HIV.

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Section: Discussionmentioning

confidence: 80%

HIV-1 Nef Targets HDAC6 to Assure Viral Production and Virus Infection

et al. 2019

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“…These studies also suggested the most important difference between the infection of these two cells was the superior ability of aCD4 T cells to release virions and initiate spreading infection (16). Multiple cellular factors have been suggested to restrict HIV infection in rCD4 T cells at multiple steps, including SAMHD1 for reverse transcription (17)(18)(19) and TRABD2A for virion release (20).…”

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confidence: 99%

Next-Generation Sequencing in a Direct Model of HIV Infection Reveals Important Parallels to and Differences from In Vivo Reservoir Dynamics

Pinzone

Bertuccio

VanBelzen

et al. 2020

J Virol

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Next-generation sequencing (NGS) represents a powerful tool to unravel the genetic make-up of the HIV reservoir, but limited data exist on its use in vitro. Moreover, most NGS studies do not separate integrated from unintegrated DNA, even though selection pressures on these two forms should be distinct. We reasoned we could use NGS to compare the infection of resting and activated CD4 T cells in vitro to address how the metabolic state affects reservoir formation and dynamics. To address these questions, we obtained HIV sequences 2, 4, and 8 days after NL4-3 infection of metabolically activated and quiescent CD4 T cells (cultured with 2 ng/ml interleukin-7). We compared the composition of integrated and total HIV DNA by isolating integrated HIV DNA using pulsed-field electrophoresis before performing sequencing. After a single-round infection, the majority of integrated HIV DNA was intact in both resting and activated T cells. The decay of integrated intact proviruses was rapid and similar in both quiescent and activated T cells. Defective forms accumulated relative to intact ones analogously to what is observed in vivo. Massively deleted viral sequences formed more frequently in resting cells, likely due to lower deoxynucleoside triphosphate (dNTP) levels and the presence of multiple restriction factors. To our surprise, the majority of these deleted sequences did not integrate into the human genome. The use of NGS to study reservoir dynamics in vitro provides a model that recapitulates important aspects of reservoir dynamics. Moreover, separating integrated from unintegrated HIV DNA is important in some clinical settings to properly study selection pressures. IMPORTANCE The major implication of our work is that the decay of intact proviruses in vitro is extremely rapid, perhaps as a result of enhanced expression. Gaining a better understanding of why intact proviruses decay faster in vitro might help the field identify strategies to purge the reservoir in vivo. When used wisely, in vitro models are a powerful tool to study the selective pressures shaping the viral landscape. Our finding that massively deleted sequences rarely succeed in integrating has several ramifications. It demonstrates that the total HIV DNA can differ substantially in character from the integrated HIV DNA under certain circumstances. The presence of unintegrated HIV DNA has the potential to obscure selection pressures and confound the interpretation of clinical studies, especially in the case of trials involving treatment interruptions.

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“…HeLa (ATCC, Cat.CCL-2), 293T (ATCC, Cat.CRL-11268), mouse NIH3T3 cells (NIH AIDS Reagent Program, Cat.ARP-9946), TZM-bl (provided by Dr Guangxia Gao) 41 , and Jurkat (ATCC,Cat.TIB-152) cells were grown and maintained in Dulbecco’s modified Eagle’s medium (Gibco) or RPMI-1640 medium (Gibco), as previously described 42 . Both media were supplemented with 10% fetal bovine serum (FBS, Gibco), 100 U/mL penicillin, and 100 mg/mL streptomycin.…”

Section: Methodsmentioning

confidence: 99%

“…To stimulate CD4 + T cells, CD3/CD28 activator magnetic beads (Invitrogen) were added to the culture medium and incubated for 2 days along with IL-2 (50 U/mL; Biomol) according to the manufacturer’s instructions. The isolation and culture of monocytes, MDMs, and MDDCs was performed as previously described 42 . Briefly, MDMs were generated by stimulating monocytes with 50 ng/mL recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF; R&D) for 7 days.…”

Section: Methodsmentioning

confidence: 99%

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Vpr counteracts the restriction of LAPTM5 to promote HIV-1 infection in macrophages

Zhao

Wang

et al. 2021

Nat Commun

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The HIV-1 accessory proteins Vif, Vpu, and Nef can promote infection by overcoming the inhibitory effects of the host cell restriction factors APOBEC3G, Tetherin, and SERINC5, respectively. However, how the HIV-1 accessory protein Vpr enhances infection in macrophages but not in CD4+ T cells remains elusive. Here, we report that Vpr counteracts lysosomal-associated transmembrane protein 5 (LAPTM5), a potent inhibitor of HIV-1 particle infectivity, to enhance HIV-1 infection in macrophages. LAPTM5 transports HIV-1 envelope glycoproteins to lysosomes for degradation, thereby inhibiting virion infectivity. Vpr counteracts the restrictive effects of LAPTM5 by triggering its degradation via DCAF1. In the absence of Vpr, the silencing of LAPTM5 precisely phenocopied the effect of Vpr on HIV-1 infection. In contrast, Vpr did not enhance HIV-1 infection in the absence of LAPTM5. Moreover, LAPTM5 was highly expressed in macrophages but not in CD4+ T lymphocytes. Re-expressing LAPTM5 reconstituted the Vpr-dependent promotion of HIV-1 infection in primary CD4+ T cells, as observed in macrophages. Herein, we demonstrate the molecular mechanism used by Vpr to overcome LAPTM5 restriction in macrophages, providing a potential strategy for anti-HIV/AIDS therapeutics.

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Membrane metalloprotease TRABD2A restricts HIV-1 progeny production in resting CD4+ T cells by degrading viral Gag polyprotein

Cited by 19 publications

References 41 publications

HIV-1 Nef Targets HDAC6 to Assure Viral Production and Virus Infection

HIV-1 Nef Targets HDAC6 to Assure Viral Production and Virus Infection

Next-Generation Sequencing in a Direct Model of HIV Infection Reveals Important Parallels to and Differences from In Vivo Reservoir Dynamics

Vpr counteracts the restriction of LAPTM5 to promote HIV-1 infection in macrophages

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