Viral Ubiquitin and Ubiquitin-Like Deconjugases—Swiss Army Knives for Infection

Masucci, Maria G.

doi:10.3390/biom10081137

Cited by 10 publications

(7 citation statements)

References 186 publications

(244 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…BPLF1 is the EBV encoded member of a family of viral ubiquitin- and NEDD8-specific deconjugases that is conserved in all human and animal herpesviruses. The enzymes play multiple roles in infection by regulating the activity and turnover of proteins that control key cellular functions required for virus replication and infectivity as well as innate antiviral defenses [ 43 ]. By using an unbiased approach based on co-immunoprecipitation with the N-terminal catalytic domain of BPLF1, we have identified several components of the vesicular trafficking and autophagic machinery as potential interacting partners of BPLF1 ( Figure 1 ).…”

Section: Discussionmentioning

confidence: 99%

The Epstein-Barr virus deubiquitinase BPLF1 targets SQSTM1/p62 to inhibit selective autophagy

2021

View full text Add to dashboard Cite

Macroautophagy/autophagy plays an important role in the control of viral infections and viruses have evolved multiple strategies to interfere with autophagy to avoid destruction and promote their own replication and spread. Here we report that the deubiquitinase encoded in the N-terminal domain of the Epstein-Barr virus (EBV) large tegument protein, BPLF1, regulates selective autophagy. Mass spectrometry analysis identified several vesicular traffic and autophagy related proteins as BPLF1 interactors and potential substrates, suggesting that the viral protein targets this cellular defense during productive infection. Direct binding of BPLF1 to the autophagy receptor SQSTM1/p62 (sequestosome 1) was confirmed by co-immunoprecipitation of transfected BPLF1 and by in vitro affinity isolation of bacterially expressed proteins. Expression of the catalytically active BPLF1 was associated with decreased SQSTM1/p62 ubiquitination and failure to recruit LC3 to SQSTM1/p62-positive aggregates. Selective autophagy was inhibited as illustrated by the accumulation of large protein aggregates in BPLF1-positive cells co-transfected with an aggregate-prone HTT (huntingtin)-Q109 construct, and by a slower autophagy-dependent clearance of protein aggregates upon transfection of BPLF1 in cells expressing a tetracycline-regulated HTT-Q103. The inhibition of aggregate clearance was restored by overexpression of a SQSTM1/p62[E409A,K420R] mutant that does not require ubiquitination of Lys420 for cargo loading. These findings highlight a previously unrecognized role of the viral deubiquitinase in the regulation of selective autophagy, which may promote infection and the production of infectious virus.

show abstract

Section: Discussionmentioning

confidence: 99%

The Epstein-Barr virus deubiquitinase BPLF1 targets SQSTM1/p62 to inhibit selective autophagy

2021

View full text Add to dashboard Cite

show abstract

“…An important role in signal transduction and regulation of the antiviral response plays the abundant ISG15 effector, which was 1.8-fold induced after SARS-CoV-2 infection in the proteome. ISG15 functions amongst others as Ubiquitin-like modifier in ISGylation of RIG-I and IRF-3, which are targeted for degradation or activated to regulate IFN and ISG production (Figure 7A) (Masucci, 2020). Widespread ISGylation of newly synthesized viral proteins is proposed to inhibit viral replication and translation (Durfee et al, 2010).…”

Section: Changes In the Calu-3 Proteome After Sars-cov-2 Infectionmentioning

confidence: 99%

The Effect of Allicin on the Proteome of SARS-CoV-2 Infected Calu-3 Cells

et al. 2021

View full text Add to dashboard Cite

Allicin (diallyl thiosulfinate) is the major thiol-reactive organosulfur compound produced by garlic plants (Allium sativum) upon tissue damage. Allicin exerts its strong antimicrobial activity against bacteria and fungi via S-thioallylation of protein thiols and low molecular weight thiols. Here, we investigated the effect of allicin on SARS-CoV-2 infected Vero E6 and Calu-3 cells. Toxicity tests revealed that Calu-3 cells showed greater allicin tolerance, probably due to >4-fold higher GSH levels compared to the very sensitive Vero E6 cells. Exposure of infected Vero E6 and Calu-3 cells to biocompatible allicin doses led to a ∼60–70% decrease of viral RNA and infectious viral particles. Label-free quantitative proteomics was used to investigate the changes in the Calu-3 proteome after SARS-CoV-2 infection and the effect of allicin on the host-virus proteome. SARS-CoV-2 infection of Calu-3 cells caused a strong induction of the antiviral interferon-stimulated gene (ISG) signature, including several antiviral effectors, such as cGAS, Mx1, IFIT, IFIH, IFI16, IFI44, OAS, and ISG15, pathways of vesicular transport, tight junctions (KIF5A/B/C, OSBPL2, CLTCL1, and ARHGAP17) and ubiquitin modification (UBE2L3/5), as well as reprogramming of host metabolism, transcription and translation. Allicin treatment of infected Calu-3 cells reduced the expression of IFN signaling pathways and ISG effectors and reverted several host pathways to levels of uninfected cells. Allicin further reduced the abundance of the structural viral proteins N, M, S and ORF3 in the host-virus proteome. In conclusion, our data demonstrate the antiviral and immunomodulatory activity of biocompatible doses of allicin in SARS-CoV-2-infected cell cultures. Future drug research should be directed to exploit the thiol-reactivity of allicin derivatives with increased stability and lower human cell toxicity as antiviral lead compounds.

show abstract

“…Ubiquitin-specific proteases, or deubiquitinating enzymes (DUBs), regulate protein turnover by disassembling poly-ubiquitin chains that target the substrate for proteasomal degradation [ 27 ]. Several human and animal viruses encode DUB homologs that play important roles in the virus life cycle by promoting viral genome replication and inhibiting the host antiviral response [ 28 – 31 ]. In this study, we report that TOP2 is a substrate of the DUB encoded in the N-terminal domain of the EBV large tegument protein BPLF1 and provide evidence for the capacity of the viral enzyme to promote the non-proteolytic TDP2-dependent resolution of TOP2ccs, which enhances cell survival and favors virus production.…”

Section: Introductionmentioning

confidence: 99%

The Epstein-Barr virus deubiquitinating enzyme BPLF1 regulates the activity of topoisomerase II during productive infection

Nagy

Liu

et al. 2021

PLoS Pathog

View full text Add to dashboard Cite

Topoisomerases are essential for the replication of herpesviruses but the mechanisms by which the viruses hijack the cellular enzymes are largely unknown. We found that topoisomerase-II (TOP2) is a substrate of the Epstein-Barr virus (EBV) ubiquitin deconjugase BPLF1. BPLF1 co-immunoprecipitated and deubiquitinated TOP2, and stabilized SUMOylated TOP2 trapped in cleavage complexes (TOP2cc), which halted the DNA damage response to TOP2-induced double strand DNA breaks and promoted cell survival. Induction of the productive virus cycle in epithelial and lymphoid cell line carrying recombinant EBV encoding the active enzyme was accompanied by TOP2 deubiquitination, accumulation of TOP2ccs and resistance to Etoposide toxicity. The protective effect of BPLF1 was dependent on the expression of tyrosyl-DNA phosphodiesterase 2 (TDP2) that releases DNA-trapped TOP2 and promotes error-free DNA repair. These findings highlight a previously unrecognized function of BPLF1 in supporting a non-proteolytic pathway for TOP2cc debulking that favors cell survival and virus production.

show abstract

Viral Ubiquitin and Ubiquitin-Like Deconjugases—Swiss Army Knives for Infection

Cited by 10 publications

References 186 publications

The Epstein-Barr virus deubiquitinase BPLF1 targets SQSTM1/p62 to inhibit selective autophagy

The Epstein-Barr virus deubiquitinase BPLF1 targets SQSTM1/p62 to inhibit selective autophagy

The Effect of Allicin on the Proteome of SARS-CoV-2 Infected Calu-3 Cells

The Epstein-Barr virus deubiquitinating enzyme BPLF1 regulates the activity of topoisomerase II during productive infection

Contact Info

Product

Resources

About