Programmed ribosomal frameshifting (PRF) is a fundamental gene expression event in many viruses, including SARS-CoV-2. It allows production of essential viral, structural and replicative enzymes that are encoded in an alternative reading frame. Despite the importance of PRF for the viral life cycle, it is still largely unknown how and to what extent cellular factors alter mechanical properties of frameshift elements and thereby impact virulence. This prompted us to comprehensively dissect the interplay between the SARS-CoV-2 frameshift element and the host proteome. We reveal that the short isoform of the zinc-finger antiviral protein (ZAP-S) is a direct regulator of PRF in SARS-CoV-2 infected cells. ZAP-S overexpression strongly impairs frameshifting and inhibits viral replication. Using in vitro ensemble and single-molecule techniques, we further demonstrate that ZAP-S directly interacts with the SARS-CoV-2 RNA and interferes with the folding of the frameshift RNA element. Together, these data identify ZAP-S as a host-encoded inhibitor of SARS-CoV-2 frameshifting and expand our understanding of RNA-based gene regulation.

The effect of liquid isoprene rubber (LIR) on the dynamic mechanical properties of emulsion‐polymerized styrene/butadiene rubber (ESBR) vulcanizates was investigated by temperature sweep using dynamic mechanical analysis. The introduction of LIR led to ESBR vulcanizates having higher loss factor (tan δ) in the temperature range − 30 to 0 °C, and lower tan δ in the range 60 to 80 °C. A small amount of LIR‐403 (LIR with carboxyl groups) led to a significant change in tan δ: the addition of LIR‐403 (3 phr) led to a 7.5% increase in tan δ from − 30 to 0 °C, and a 24.9% decrease in tan δ from 60 to 80 °C. It was found that the introduction of LIR increased the bound rubber content in the ESBR compound. Equilibrium swelling experiments showed that the crosslink density of the vulcanizates increased after the introduction of LIR‐403 or LIR‐50 (general purpose LIR). The change in tan δ from 60 to 80 °C was related to polymer–filler interactions. The characteristic constant of filler–ESBR matrix interaction (m) was calculated. At a given filler volume fraction, the increase in m in the presence of LIR could be well related to the decrease in tan δ from 60 to 80 °C. The influence of LIR on filler network in the ESBR compound was also investigated by strain and temperature sweeps using a rubber processing analyzer. Copyright © 2011 Society of Chemical Industry

Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. HCV can be sensed by host innate immunity to induce expression of interferons (IFNs) and a number of antiviral effectors. In this study, we found HCV infection induced the expression of Neuralized E3 Ubiquitin Protein Ligase 3 (NEURL3), a putative E3 ligase, in a manner that requires the involvement of innate immune sensing but is independent of the IFN action. Furthermore, we showed that NEURL3 inhibited HCV infection, while had little effect on other RNA viruses including zika virus, dengue virus and vesicular stomatitis virus. Mechanistic studies demonstrated that NEURL3 inhibited HCV assembly by directly binding HCV envelope glycoprotein E1 to interfere with the E1/E2 heterodimerization, an important prerequisite for virion morphogenesis. Finally, we showed that knockout of NEURL3 significantly enhanced HCV infection. In summary, we identified NEURL3 as a novel inducible antiviral host factor that suppresses HCV assembly. Our results not only shed new insight into how host innate immunity acts against HCV, but also revealed a new important biological function for NEURL3. The exact biological function of NEURL3, a putative E3 ligase remains largely unknown. In this study, we found that NEURL3 could be upregulated upon HCV infection in a manner dependent on pattern-recognition receptor-mediated innate immune response. NEURL3 inhibits HCV assembly by directly binding viral E1 envelope glycoprotein to disrupt its interaction with E2, an action that requires its NHR domain but not RING domain. Furthermore, we found that NEURL3 has a pan-genotypic anti-HCV activity and interacts with E1 of genotype 2a, 1b, 3a and 6a, but does not inhibit other closely related RNA viruses such as ZIKV, DENV and VSV. To our knowledge, our study is the first report to demonstrate that NEURL3 functions as an antiviral host factor. Our results not only shed new insight into how host innate immunity acts against HCV, but also revealed a new important biological function for NEURL3.