Inhibitors of the Ubiquitin-Mediated Signaling Pathway Exhibit Broad-Spectrum Antiviral Activities against New World Alphaviruses

Boghdeh, Niloufar; McGraw, Brittany; Barrera, Michael D; Anderson, Carol; Baha, Haseebullah; Risner, Kenneth; Ogungbe, Ifedayo Victor; Alem, Farhang; Narayanan, Aarthi

doi:10.3390/v15030655

Cited by 6 publications

(4 citation statements)

References 85 publications

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“…We also show that E3 ligase activity is required for TRIM32-mediated inhibition of VEEV. However, we could not assess whether UPS inhibitors can subvert TRIM32-mediated inhibition of VEEV, because the host UPS also required for virus infection [ 19 , 21 ]. Previous study reported that TRIM32 mutants R394H and D487N lost monoubiquitination feature, but still remain the ability to self-associate.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies found that the host ubiquitin proteasome system (UPS) plays a crucial role in alphaviruses infection. Specifically, the post-entry stages of alphaviruses can be disrupted by UPS inhibitors, such as proteasome inhibitors and deubiquitinating enzyme inhibitors [19][20][21]. To date, only a few E3 ubiquitin ligases (E3) and deubiquitinases (DUB) have been identified as host factor or restriction factor for alphaviruses.…”

Section: Introductionmentioning

confidence: 99%

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TRIM32 inhibits Venezuelan Equine Encephalitis Virus Infection by targeting a late step in viral entry

Xie,

Cao,

Gan

et al. 2024

Preprint

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Alphaviruses are mosquito borne RNA viruses that are a reemerging public health threat. Alphaviruses have a broad host range, and can cause diverse disease outcomes like arthritis, and encephalitis. The host ubiquitin proteasome system (UPS) plays critical roles in regulating cellular processes to control the infections with various viruses, including alphaviruses. Previous studies suggest alphaviruses hijack UPS for virus infection, but the molecular mechanisms remain poorly characterized. In addition, whether certain E3 ubiquitin ligases or deubiquitinases act as alphavirus restriction factors remains poorly understood. Here, we employed a cDNA expression screen to identify E3 ubiquitin ligase TRIM32 as a novel intrinsic restriction factor against alphavirus infection, including VEEV-TC83, SINV, and ONNV. Ectopic expression of TRIM32 reduces alphavirus infection, whereas depletion of TRIM32 with CRISPR-Cas9 increases infection. We demonstrate that TRIM32 inhibits alphaviruses through a mechanism that is independent of the TRIM32-STING-IFN axis. Combining reverse genetics and biochemical assays, we found that TRIM32 interferes with genome translation after membrane fusion, prior to replication of the incoming viral genome. Furthermore, our data indicate that the monoubiquitination of TRIM32 is important for its antiviral activity. Notably, we also show two TRIM32 pathogenic mutants R394H and D487N, related to Limb-girdle muscular dystrophy (LGMD), have a loss of antiviral activity against VEEV-TC83. Collectively, these results reveal that TRIM32 acts as a novel intrinsic restriction factor suppressing alphavirus infection and provides insights into the interaction between alphaviruses and the host UPS.Author summaryDue to climate change, wildlife habitat loss, and human activities, alphavirus infections are a growing threat to public health. The host UPS has critical role in virus-host interaction, but how the UPS impact alphavirus infection is not completely understood. In this study, we found that the E3 ubiquitin ligase TRIM32 inhibits diverse alphaviruses in multiple cell types. Mechanistically, TRIM32 impairs primary translation of incoming viral genome in a manner that depends on. monoubiquitination of TRIM32. Additionally, disease-associated alleles of TRIM32 have a loss-of-function with respect to viral inhibition. Together, these findings uncover a novel biological function of TRIM32 in regulating alphavirus infection and provide important insights into the interplay between alphaviruses and the host USP.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

TRIM32 inhibits Venezuelan Equine Encephalitis Virus Infection by targeting a late step in viral entry

Xie,

Cao,

Gan

et al. 2024

Preprint

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“…This compound also possessed the highest cytotoxicity among the examined compounds (Figure 2). This compound may have further application prospects due to its demonstrated antitumor and antiviral activity, including the inhibition of the growth of implanted human tumors without causing toxic effects in mice [26,41,42].…”

Section: Discussionmentioning

confidence: 99%

Enzymatic Redox Properties and Cytotoxicity of Irreversible Nitroaromatic Thioredoxin Reductase Inhibitors in Mammalian Cells

Nemeikaitė-Čėnienė,

Misevičienė,

Marozienė

et al. 2023

IJMS

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NADPH:thioredoxin reductase (TrxR) is considered a potential target for anticancer agents. Several nitroheterocyclic sulfones, such as Stattic and Tri-1, irreversibly inhibit TrxR, which presumably accounts for their antitumor activity. However, it is necessary to distinguish the roles of enzymatic redox cycling, an inherent property of nitroaromatics (ArNO2), and the inhibition of TrxR in their cytotoxicity. In this study, we calculated the previously unavailable values of single-electron reduction potentials of known inhibitors of TrxR (Stattic, Tri-1, and 1-chloro-2,4-dinitrobenzene (CDNB)) and inhibitors identified (nitrofuran NSC697923 and nitrobenzene BTB06584). These calculations were according to the rates of their enzymatic single-electron reduction (PMID: 34098820). This enabled us to compare their cytotoxicity with that of model redox cycling ArNO2. In MH22a and HCT-116 cells, Tri-1, Stattic, CDNB, and NSC697023 possessed at least 10-fold greater cytotoxicity than can be expected from their redox cycling activity. This may be related to TrxR inhibition. The absence of enhanced cytotoxicity in BTB06548 may be attributed to its instability. Another known inhibitor of TrxR, tetryl, also did not possess enhanced cytotoxicity, probably because of its detoxification by DT-diaphorase (NQO1). Apart from the reactions with NQO1, the additional mechanisms influencing the cytotoxicity of the examined inhibitors of TrxR are their reactions with cytochromes P-450. Furthermore, some inhibitors, such as Stattic and NSC697923, may also inhibit glutathione reductase. We suggest that these data may be instrumental in the search for TrxR inhibitors with enhanced cytotoxic/anticancer activity.

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“…Negative-strand synthesis was performed as previously described [23]. cDNA was generated using a specific primer to negative-strand RNA for VEEV TC-83, which contained a T7 promoter sequence attached at the 5 ′ end (T7-TC83-Neg 5 ′ -GCGTAATACGACTCACTAT ATCCGTCAGCTCTCTCGCAGG-3 ′ ).…”

Section: Negative-strand Rt-qpcrmentioning

confidence: 99%

Brilacidin as a Broad-Spectrum Inhibitor of Enveloped, Acutely Infectious Viruses

Anderson,

Barrera,

Boghdeh

et al. 2023

Microorganisms

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Alphaviruses, belonging to the Togaviridae family, and bunyaviruses, belonging to the Paramyxoviridae family, are globally distributed and lack FDA-approved vaccines and therapeutics. The alphaviruses Venezuelan equine encephalitis virus (VEEV) and eastern equine encephalitis virus (EEEV) are known to cause severe encephalitis, whereas Sindbis virus (SINV) causes arthralgia potentially persisting for years after initial infection. The bunyavirus Rift Valley Fever virus (RVFV) can lead to blindness, liver failure, and hemorrhagic fever. Brilacidin, a small molecule that was designed de novo based on naturally occurring host defensins, was investigated for its antiviral activity against these viruses in human small airway epithelial cells (HSAECs) and African green monkey kidney cells (Veros). This testing was further expanded into a non-enveloped Echovirus, a Picornavirus, to further demonstrate brilacidin’s effect on early steps of the viral infectious cycle that leads to inhibition of viral load. Brilacidin demonstrated antiviral activity against alphaviruses VEEV TC-83, VEEV TrD, SINV, EEEV, and bunyavirus RVFV. The inhibitory potential of brilacidin against the viruses tested in this study was dependent on the dosing strategy which necessitated compound addition pre- and post-infection, with addition only at the post-infection stage not eliciting a robust inhibitory response. The inhibitory activity of brilacidin was only modest in the context of the non-enveloped Picornavirus Echovirus, suggesting brilacidin may be less potent against non-enveloped viruses.

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Inhibitors of the Ubiquitin-Mediated Signaling Pathway Exhibit Broad-Spectrum Antiviral Activities against New World Alphaviruses

Cited by 6 publications

References 85 publications

TRIM32 inhibits Venezuelan Equine Encephalitis Virus Infection by targeting a late step in viral entry

TRIM32 inhibits Venezuelan Equine Encephalitis Virus Infection by targeting a late step in viral entry

Enzymatic Redox Properties and Cytotoxicity of Irreversible Nitroaromatic Thioredoxin Reductase Inhibitors in Mammalian Cells

Brilacidin as a Broad-Spectrum Inhibitor of Enveloped, Acutely Infectious Viruses

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