Revealing the host antiviral protein ZAP-S as an inhibitor of SARS-CoV-2 programmed ribosomal frameshifting

Zimmer, Matthias M.; Kibe, Anuja; Rand, Ulfert; Pekarek, Lukas; Čičin‐Šain, Luka; Caliskan, Neva

doi:10.1101/2021.05.31.445667

Cited by 4 publications

(4 citation statements)

References 80 publications

(114 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, while essential for RNA-binding and CpG specificity, the RBD likely has no intrinsic antiviral activity alone at physiological expression levels. A recent preprint has suggested that ZAP-S can inhibit SARS CoV-2 by negatively regulating the -1 frameshift between Orf1a and Orf1b [48]. Consistent with this, we do see a small reduction in N expression in infected cells expressing ZAP-S alone, but it is insufficient to significantly impact viral production in the supernatant.…”

Section: Discussionsupporting

confidence: 88%

The C-terminal PARP domain of the long ZAP isoform contributes essential effector functions for CpG-directed antiviral activity

Kmieć

Lista-Brotos

Ficarelli

et al. 2021

Preprint

View full text Add to dashboard Cite

The zinc finger antiviral protein (ZAP) is a broad inhibitor of virus replication. Its best-characterized function is to bind CpG dinucleotides present in viral RNA and, through the recruitment of TRIM25, KHNYN and other cellular RNA degradation machinery, target them for degradation or prevent their translation. ZAP’s activity requires the N-terminal RNA binding domain that selectively binds CpG-containing RNA. However, much less is known about the functional contribution of the remaining domains. Using ZAP-sensitive and ZAP-insensitive human immunodeficiency virus type I (HIV-1), we show that the catalytically inactive poly-ADP-ribose polymerase (PARP) domain of the long ZAP isoform (ZAP-L) is essential for CpG-specific viral restriction. Mutation of a crucial cysteine in the C-terminal CaaX box that mediates S-farnesylation and, to a lesser extent, the inactive catalytic site triad within the PARP domain, disrupted the activity of ZAP-L. Addition of the CaaX box to ZAP-S partly restored antiviral activity, explaining why ZAP-S lacks CpG-dependent antiviral activity despite conservation of the RNA-binding domain. Confocal microscopy confirmed the CaaX motif mediated localization of ZAP-L to vesicular structures and enhanced physical association with intracellular membranes. Importantly, the PARP domain and CaaX box together modulate the interaction between ZAP-L and its cofactors TRIM25 and KHNYN, implying that its proper subcellular localisation is required to establish an antiviral complex. The essential contribution of the PARP domain and CaaX box to ZAP-L’s CpG-directed antiviral activity was further confirmed by inhibition of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication. Thus, compartmentalization of ZAP-L on intracellular membranes provides an essential effector function in the ZAP-L-mediated antiviral activity.

show abstract

Section: Discussionsupporting

confidence: 88%

The C-terminal PARP domain of the long ZAP isoform contributes essential effector functions for CpG-directed antiviral activity

Kmieć

Lista-Brotos

Ficarelli

et al. 2021

Preprint

View full text Add to dashboard Cite

show abstract

“…In epithelial cell lines, ATXN2L regulates stress granules and processing bodies (Kaehler et al, 2012). ZC3HAV1 was shown to destabilize IFNB and IFNL3 mRNAs in hepatocytes, and human cytomegalovirus RNA in fibroblasts, and it inhibits programmed ribosomal frameshifting of the SARS-CoV-2 virus (Schwerk et al, 2019;Gonzalez-Perez et al, 2021;Zimmer et al, 2021). The mechanisms that ATXN2L and ZC3HAV1 employ in T cells to supress cytokine production, however, await further elucidation.…”

Section: Discussionmentioning

confidence: 99%

Time-dependent regulation of cytokine production by RNA binding proteins defines T cell effector function

Popović

Guislain

Engels

et al. 2021

Preprint

View full text Add to dashboard Cite

SummaryEffective T cell responses against target cells require controlled production of the key pro-inflammatory cytokines IFN-γ, TNF and IL-2. Post-transcriptional events determine the magnitude and duration of cytokine production in T cells, a process that is largely regulated by RNA binding proteins (RBPs). Here we studied the identity and mode of action of RBPs interacting with cytokine mRNAs. With an RNA aptamer-based capture assay from human T cell lysates, we identified >130 RBPs interacting with the full length 3’untranslated regions of IFNG, TNF and IL2. The RBP landscape altered upon T cell activation. Furthermore, RBPs display temporal activity profiles to control cytokine production. Whereas HuR promotes early cytokine production, the peak production levels and response duration is controlled by ZFP36L1, ATXN2L and ZC3HAV1. Importantly, ZFP36L1 deletion boosts T cell responses against tumors in vivo, revealing the potential of the RBP map to identify critical modulators of T cell responses.

show abstract

“…Another important antiviral mechanism is the inhibition of protein translation from viral mRNAs. The binding of PARP13 to viral RNAs can reduce their association with ribosomes, reducing their translation [45,46], and recent evidence suggests that specific ribosome frameshifting events, such as the ORF1a/b slippage event in SARS-CoV-2 can be regulated by PARP13 binding as well [47]. Strong inhibition of protein synthesis was also observed in cells overexpressing PARP7, PARP10 or PARP12L (the long isoform of PARP12) from virally encoded constructs, although a clear mechanistic understanding of this effect is currently lacking [28,29,48].…”

Section: Adp-ribosylation and Ifn-induced Effector Functionsmentioning

confidence: 99%

Host ADP-ribosylation and the SARS-CoV-2 macrodomain

Hoch

2021

Biochemical Society Transactions

View full text Add to dashboard Cite

The COVID-19 pandemic has prompted intense research efforts into elucidating mechanisms of coronavirus pathogenesis and to propose antiviral interventions. The interferon (IFN) response is the main antiviral component of human innate immunity and is actively suppressed by several non-structural SARS-CoV-2 proteins, allowing viral replication within human cells. Differences in IFN signalling efficiency and timing have emerged as central determinants of the variability of COVID-19 disease severity between patients, highlighting the need for an improved understanding of host–pathogen interactions that affect the IFN response. ADP-ribosylation is an underexplored post-translational modification catalyzed by ADP-ribosyl transferases collectively termed poly(ADP-ribose) polymerases (PARPs). Several human PARPs are induced by the IFN response and participate in antiviral defences by regulating IFN signalling itself, modulating host processes such as translation and protein trafficking, as well as directly modifying and inhibiting viral target proteins. SARS-CoV-2 and other viruses encode a macrodomain that hydrolyzes ADP-ribose modifications, thus counteracting antiviral PARP activity. This mini-review provides a brief overview of the known targets of IFN-induced ADP-ribosylation and the functions of viral macrodomains, highlighting several open questions in the field.

show abstract

Revealing the host antiviral protein ZAP-S as an inhibitor of SARS-CoV-2 programmed ribosomal frameshifting

Cited by 4 publications

References 80 publications

The C-terminal PARP domain of the long ZAP isoform contributes essential effector functions for CpG-directed antiviral activity

The C-terminal PARP domain of the long ZAP isoform contributes essential effector functions for CpG-directed antiviral activity

Time-dependent regulation of cytokine production by RNA binding proteins defines T cell effector function

Host ADP-ribosylation and the SARS-CoV-2 macrodomain

Contact Info

Product

Resources

About