Analysis of individual HIV-1 budding event using fast AFM reveals a multiplexed role for VPS4

Harel, Shimon; Altaras, Yarin; Nachmias, Dikla; Rotem-Dai, Noa; Dvilansky, Inbar; Elia, Natalie; Rousso, Itay

doi:10.1016/j.bpj.2022.08.035

Cited by 10 publications

(16 citation statements)

References 45 publications

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“…Nevertheless, depleting VPS4A in cancer cells with loss of VPS4B led to synthetic cell death, suggesting at least partial overlap between these isoforms ( 8 ). Third, VPSB was previously shown to be more crucial for HIV viral release than VPS4A, and we recently substantiated these findings and showed that depletion of VPS4A or VPS4B has differential effects on HIV-viral release, suggesting a unique property for each isoform ( 14, 15 ). Collectively, these reports raise possibility that the two VPS4 isoforms hold overlapping and unique functions in cells stressing the need to investigate the functional differences of these two isoforms in cells.…”

Section: Introductionsupporting

confidence: 60%

“…To dissect the role of VPS4 isoforms in cytokinesis, we monitored abscission in VPSA and VPS4B KO cells recently generated in our laboratory (Fig. S1A) ( 14 ). Attempts to generate cells depleted of both VPS4A and VPS4B resulted in cell death, suggesting that at least one VPS4 isoform is needed for cell viability, as previously suggested ( 8 ).…”

Section: Resultsmentioning

confidence: 99%

“…VTA1 is a co-factor of VPS4 that was shown to stabilize its active, hexameric form ( 11, 12, 26-28 ). To test whether the regulatory role of VPS4 can be induced by destabilizing the VPS4 hexamer, we examined the effect of VTA1 depletion on VPS4 function in dividing cells using a VTA1 KO cell line ( 14 ) (Fig. S2A).…”

Section: Resultsmentioning

confidence: 99%

“…The role of VPS4 in the ESCRT pathway has been studied both in vitro and in cells. Perturbation of VPS4 function inhibited ESCRT-mediated processes in cells, including viral budding and cytokinetic abscission ( 3, 14–16, 21, 22, 29, 30 ). In vitro, VPS4 was induced the disassembly of ESCRT-III filaments and to drive ESCRT-mediated membrane fission in artificial membrane vesicles, indicating that it is executing the final membrane fission event by depolymerization ESCRT-III filaments ( 16, 31-36 ).…”

Section: Discussionmentioning

confidence: 99%

“…Differences in VPS4A and VPS4B function in cells have been previously reported in the context of viral budding. In particular, it was shown that the HIV virus mainly utilizes the VPS4B isoform to bud out of cells ( 14, 15 ). Our observations that the cellular levels of VPS4B are significantly higher than VPS4A and that VPS4A is involved in the regulation of abscission may provide a logical explanation for these observations.…”

Section: Discussionmentioning

confidence: 99%

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Differential role for VPS4 isoforms in cytokinetic abscission confers a regulatory function for monomeric VPS4A and VTA1 in mammalian cells

Dvilansky,

Atlaras,

Nachmias

et al. 2023

Preprint

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Mutations in the human AAA-ATPase VPS4 isoform, VPS4A, cause severe Neurodevelopmental defects and Congenital Dyserythropoietic Anemia (CDA). VPS4 is a crucial component of the ESCRT system, which drives membrane remodeling in numerous cellular processes, including receptor degradation, cell division, and neural pruning. Notably, while most organisms encode for a single VPS4 gene, human cells have two VPS4 paralogs, namely VPS4A and VPS4B, but the functional differences between these paralogs is mostly unknown. Here, we set out to investigate the role of the human VPS4 paralogs in cytokinetic abscission using a series of knockout cell lines. We found that VPS4A and VPS4B hold both overlapping and distinct roles in abscission. VPS4A depletion resulted in a severe abscission delay, which was fully rescued by VPS4A expression but only partially rescued by VPS4B overexpression. Unexpectedly, expressing a monomeric-locked VPS4A mutant also partially rescued the abscission delay in VPS4A KO cells and bound the abscission checkpoint proteins CHMP4C and ANCHR. Depletion of VTA1, a co-factor of VPS4, disrupted VPS4A- ANCHR interactions and accelerated abscission, indicating a role for VTA1 in the abscission checkpoint. Our findings reveal a dual role for VPS4A in abscission, one that is canonical and can be compensated by VPS4B, and another that is regulatory and is mediated by its monomeric form. These observations provide a potential mechanistic explanation for the neurodevelopmental defects and other related disorders reported in VPS4A-mutated patients with a fully functional VPS4B paralog.

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Section: Introductionsupporting

confidence: 60%

Section: Resultsmentioning

confidence: 99%