HIV-1 Vpr-induced cell death in Schizosaccharomyces pombe is reminiscent of apoptosis

Huard, Sylvain; Chen, Mingzhong; Burdette, Kristen; Fenyvuesvolgyi, Csaba; Yu, Min; Elder, Robert E.; Zhao, Yuqi

doi:10.1038/cr.2008.272

Cited by 25 publications

(47 citation statements)

References 76 publications

(165 reference statements)

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“…Morphology of fission yeast mitochondria was visualized by staining with a mitochondria-specific dye, 2-(4-dimethylaminostyryl)-1-methylpyridinium iodide (DASPMI) [29, 36]. Consistent with previous descriptions of normal fission yeast mitochondrial morphologies [32], the mitochondria in the control cells appeared as tubular networks extending along the periphery of the cells (Fig.…”

Section: Resultssupporting

confidence: 59%

“…To explore the molecular mechanism underlying HIV-1 mdr PR-induced cell death, possible intracellular stress induced by HIV-1 mdr PRs was determined by the production of oxidative stress species (ROS) [29]. A ROS-specific dye, dihydroethidium (DHE), which produces red fluorescence in the presence of ROS, was used to measure cellular oxidative stress in the mdr PR -expressing cells.…”

Section: Resultsmentioning

confidence: 99%

“…The induction of ROS was shown by increased and visible staining of DHE. The mitochondrial morphologies ( b ) were visualized by staining of the fission yeast cells with a mitochondria-specific fluorescent probe DASPMI [29, 41]. Note that normal mitochondria typically appear like threads or necklaces constituted with multiple small dots concentrated around the edge, or as a tubular network extended along the cell peripheries.…”

Section: Resultsmentioning

confidence: 99%

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A fission yeast cell-based system for multidrug resistant HIV-1 proteases

Benkő

Dong

et al. 2017

Cell Biosci

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BackgroundHIV-1 protease (PR) is an essential enzyme for viral production. Thus, PR inhibitors (PIs) are the most effective class of anti-HIV drugs. However, the main challenge to the successful use of PI drugs in patient treatment is the emergence of multidrug resistant PRs (mdrPRs). This study aimed to develop a fission yeast cell-based system for rapid testing of new PIs that combat mdrPRs.ResultsThree mdrPRs were isolated from HIV-infected patients that carried seven (M7PR), ten (M10PR) and eleven (M11PR) PR gene mutations, respectively. They were cloned and expressed in fission yeast under an inducible promoter to allow the measurement of PR-specific proteolysis and drug resistance. The results showed that all three mdrPRs maintained their abilities to proteolyze HIV viral substrates (MA↓CA and p6) and to confer drug resistance. Production of these proteins in the fission yeast caused cell growth inhibition, oxidative stress and altered mitochondrial morphologies that led to cell death. Five investigational PIs were used to test the utility of the established yeast system with an FDA-approved PI drug Darunavir (DRV) as control. All six compounds suppressed the wildtype PR (wtPR) and the M7PR-mediated activities. However, none of them were able to suppress the M10PR or the M11PR.ConclusionsThe three clinically isolated mdrPRs maintained their viral proteolytic activities and drug resistance in the fission yeast. Furthermore, those viral mdrPR activities were coupled with the induction of growth inhibition and cell death, which could be used to test the PI activities. Indeed, the five investigational PIs and DRV suppressed the wtPR in fission yeast as they did in mammalian cells. Significantly, two of the high level mdrPRs (M10PR and M11PR) were resistant to all of the existing PI drugs including DRV. This observation underscores the importance of continued searching for new PIs against mdrPRs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13578-016-0131-5) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 59%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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A fission yeast cell-based system for multidrug resistant HIV-1 proteases

Benkő

Dong

et al. 2017

Cell Biosci

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“…Notably, Vpr also interacts with ANT and/or VDAC, and dissipates ΔΨ m (Jacotot et al, 2001; Jacotot et al, 2000). Moreover, Vpr alone is able to increase the ROS production in both mammalian cells and fission yeasts (Deshmane et al, 2009; Ferrucci et al, 2012; Huard et al, 2008; Stromajer-Racz et al, 2010). Accordingly, Vpr may also promote HIV-1 Env folding by activating the oxidative stress pathway, resulting in protection of Env from degradation via the ERAD pathway.…”

Section: Discussionmentioning

confidence: 99%

HIV-1 Vpr increases Env expression by preventing Env from endoplasmic reticulum-associated protein degradation (ERAD)

et al. 2016

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Vpr enhances HIV-1 replication in macrophages and dendritic cells, as well as the human CD4+ CEM.NKR T cell line. Recently, Vpr was reported to increase HIV-1 Env expression in macrophages. Here, we report that Vpr also increases HIV-1 Env expression in dendritic cells and CEM.NKR cells. The Vpr activity depends on its N-terminal region, which was disrupted by a single A30L mutation. Env was rapidly degraded in the absence of Vpr, which was blocked by the ERAD pathway inhibitor kifunesine or the lysosome inhibitor Bafilomycin. As2O3 or PK11195, which reportedly enhances HIV-1 Env folding, also blocked the Env degradation in CEM.NKR cells. Thus, these results not only identify Env as a primary target for Vpr to boost HIV-1 replication, but also suggest that Vpr likely promotes Env folding in the ER, which is otherwise misfolded and targeted by the ERAD pathway to lysosomes for degradation.

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“…The induction of cellular oxidative stress by ZIKV proteins was measured as the expression of ROS, which was detected by a ROS-specific dye, DHE (Sigma), that produces red fluorescence in the presence of ROS, as described previously (82,84,85). Cells were grown as described above; 48 h after GI, DHE was added to a final concentration of 5 μg/mL, and ROS were detected by fluorescent microscopy.…”

Section: Methodsmentioning

confidence: 99%

Characterization of cytopathic factors through genome-wide analysis of the Zika viral proteins in fission yeast

Poulsen

Fenyvuesvolgyi

et al. 2017

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

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The Zika virus (ZIKV) causes microcephaly and the Guillain-Barré syndrome. Little is known about how ZIKV causes these conditions or which ZIKV viral protein(s) is responsible for the associated ZIKVinduced cytopathic effects, including cell hypertrophy, growth restriction, cell-cycle dysregulation, and cell death. We used fission yeast for the rapid, global functional analysis of the ZIKV genome. All 14 proteins or small peptides were produced under an inducible promoter, and we measured the intracellular localization and the specific effects on ZIKV-associated cytopathic activities of each protein. The subcellular localization of each ZIKV protein was in overall agreement with its predicted protein structure. Five structural and two nonstructural ZIKV proteins showed various levels of cytopathic effects. The expression of these ZIKV proteins restricted cell proliferation, induced hypertrophy, or triggered cellular oxidative stress leading to cell death. The expression of premembrane protein (prM) resulted in cell-cycle G1 accumulation, whereas membrane-anchored capsid (anaC), membrane protein (M), envelope protein (E), and nonstructural protein 4A (NS4A) caused cell-cycle G2/M accumulation. A mechanistic study revealed that NS4A-induced cellular hypertrophy and growth restriction were mediated specifically through the target of rapamycin (TOR) cellular stress pathway involving Tor1 and type 2A phosphatase activator Tip41. These findings should provide a reference for future research on the prevention and treatment of ZIKV diseases. T he recent Zika virus (ZIKV) outbreak was surprising in its rapidity and alarming in its association with microcephaly in newborns (1-3) and the Guillain-Barré syndrome in adults (4, 5). Currently, little is known about ZIKV pathogenicity. ZIKV infects various neuronal cells and affects brain neurogenesis by reducing cellular proliferation and inducing cell-cycle abnormalities, cell death/apoptosis, and autophagy (2, 3, 6-8). However, it is not known which viral determinant(s) cause these cytopathic effects. The objective of this study was to identify ZIKV factors responsible for the ZIKV-mediated cytopathic effects that underlie the diseases associated with ZIKV.

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HIV-1 Vpr-induced cell death in Schizosaccharomyces pombe is reminiscent of apoptosis

Cited by 25 publications

References 76 publications

A fission yeast cell-based system for multidrug resistant HIV-1 proteases

A fission yeast cell-based system for multidrug resistant HIV-1 proteases

HIV-1 Vpr increases Env expression by preventing Env from endoplasmic reticulum-associated protein degradation (ERAD)

Characterization of cytopathic factors through genome-wide analysis of the Zika viral proteins in fission yeast

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