Chikungunya virus non-structural protein 2-mediated host shut-off disables the unfolded protein response

Fros, Jelke J.; Major, Lee; Scholte, Florine E. M.; Gardner, Joy; Hemert, Martijn J. van; Suhrbier, Andreas; Pijlman, Gorben P.

doi:10.1099/vir.0.071845-0

Cited by 62 publications

(65 citation statements)

References 53 publications

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“…Recently, the IRE-1/JNK pathway stemming from the UPR was also implicated in JEVinduced apoptosis (58). CHIKV (59) and SFV (60) induce the UPR, through the synthesis of the structural glycoproteins in the ER lumen, and this might be one possible mechanism for alphavirus-induced p38 MAPK and JNK signaling, which would not occur in the trans-replication system. Here, we show that CHIKV infection activated the p38 MAPK and JNK pathways (Fig.…”

Section: Discussionmentioning

confidence: 99%

The Antiviral Alkaloid Berberine Reduces Chikungunya Virus-Induced Mitogen-Activated Protein Kinase Signaling

Varghese

Thaa

Amrun

et al. 2016

J Virol

113

101

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Chikungunya virus (CHIKV) has infected millions of people in the tropical and subtropical regions since its reemergence in the last decade. We recently identified the nontoxic plant alkaloid berberine as an antiviral substance against CHIKV in a highthroughput screen. Here, we show that berberine is effective in multiple cell types against a variety of CHIKV strains, also at a high multiplicity of infection, consolidating the potential of berberine as an antiviral drug. We excluded any effect of this compound on virus entry or on the activity of the viral replicase. A human phosphokinase array revealed that CHIKV infection specifically activated the major mitogen-activated protein kinase (MAPK) signaling pathways extracellular signal-related kinase (ERK), p38 and c-Jun NH 2 -terminal kinase (JNK). Upon treatment with berberine, this virus-induced MAPK activation was markedly reduced. Subsequent analyses with specific inhibitors of these kinases indicated that the ERK and JNK signaling cascades are important for the generation of progeny virions. In contrast to specific MAPK inhibitors, berberine lowered virus-induced activation of all major MAPK pathways and resulted in a stronger reduction in viral titers. Further, we assessed the in vivo efficacy of berberine in a mouse model and measured a significant reduction of CHIKV-induced inflammatory disease. In summary, we demonstrate the efficacy of berberine as a drug against CHIKV and highlight the importance of the MAPK signaling pathways in the alphavirus infectious cycle. IMPORTANCEChikungunya virus (CHIKV) is a mosquito-borne virus that causes severe and persistent muscle and joint pain and has recently spread to the Americas. No licensed drug exists to counter this virus. In this study, we report that the alkaloid berberine is antiviral against different CHIKV strains and in multiple human cell lines. We demonstrate that berberine collectively reduced the virus-induced activation of cellular mitogen-activated protein kinase signaling. The relevance of these signaling cascades in the viral life cycle was emphasized by specific inhibitors of these kinase pathways, which decreased the production of progeny virions. Berberine significantly reduced CHIKV-induced inflammatory disease in a mouse model, demonstrating efficacy of the drug in vivo. Overall, this work makes a strong case for pursuing berberine as a potential anti-CHIKV therapeutic compound and for exploring the MAPK signaling pathways as antiviral targets against alphavirus infections. C hikungunya virus (CHIKV) is the causative agent of chikungunya fever, a disease spread by Aedes mosquitoes and characterized by a sudden onset of febrile illness, nausea, headache, and most importantly, severe and persistent musculoskeletal pain (1). It reemerged in tropical Asia and Africa 1 decade ago and since 2013 has infected more than 1.5 million people in the Americas (2). So far, no licensed vaccine or antiviral treatment exists to counter this disease. Over the years, many research groups have focused on...

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Section: Discussionmentioning

confidence: 99%

The Antiviral Alkaloid Berberine Reduces Chikungunya Virus-Induced Mitogen-Activated Protein Kinase Signaling

Varghese

Thaa

Amrun

et al. 2016

J Virol

113

101

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“…Total RNA was extracted from RAW 264.7 cells (1 3 10 6 ) with RNeasy Plus (Qiagen, Hilden, Germany), and 1 mg total RNA was reverse transcribed into the first-strand cDNA with ImProm (Promega, Madison WI, USA) and oligo (dT) [12][13][14][15][16][17][18] primer, according to the manufacturer's instructions. Forward 59-CCGCAGCACTCAGACTATG-39 and reverse 59-GGGTCCAACTTGTCCAGAAT-39 pairs were used to amplify uXBP1 mRNA; forward 59-CTGAGTCCGCAGCAGGT-39and reverse 59-AAACATGACAGGGTCCAACTT-39 primers were used to amplify sXBP1 mRNA; forward: 59-TCCAAGAAAGGA-CGAACATTC-39 and reverse 59-TGAGGACATCTCCCACG-TCAA-39 primers were used to amplify IFN1-b; and forward 59-GTGTCCGTCGTGGATCTG-39 and reverse 59-TGCTTCAC-CACCTTCTTGA-39 primers were used to amplify GAPDH.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

“…For example, Chikungunya virus promotes partial splicing of XBP1, although this effect does not appear to result in nuclear translocation (15). Coronavirus similarly favors expression of sXBP1 to protect infected cells from apoptosis (16).…”

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confidence: 99%

The integrated endoplasmic reticulum stress response in Leishmania amazonensis macrophage infection: the role of X‐box binding protein 1 transcription factor

et al. 2015

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Endoplasmic reticulum (ER) stress triggers the integrated ER-stress response (IERSR) that ensures cellular survival of ER stress and represents a primordial form of innate immunity. We investigated the role of IERSR during Leishmania amazonensis infection. Treatment of RAW 264.7 infected macrophages with the ER stress-inducing agent thapsigargin (TG; 1 mM) increased L. amazonensis infectivity in an IFN1-a receptor (IFNAR)-dependent manner. In Western blot assays, we showed that L. amazonensis activates the inositol-requiring enzyme (IRE1)/ X-box binding protein (XBP)-1-splicing arms of the IERSR in host cells. In chromatin immunoprecipitation (ChIP) assays, we showed an increased occupancy of enhancer and promoter sequences for the Ifnb gene by XBP1 in infected RAW 264.7 cells. Knocking down XBP1 expression by transducing RAW 264.7 cells with the short hairpin XBP1 lentiviral vector significantly reduced the parasite proliferation associated with impaired translocation of phosphorylated IFN regulatory transcription factor (IRF)-3 to the nucleus and a decrease in IFN1-b expression. Knocking down XBP1 expression also increased NO concentration, as determined by Griess reaction and reduced the expression of antioxidant genes, such as heme oxygenase (HO)-1, that protect parasites from oxidative stress. We conclude that L. amazonensis activation of XBP1 plays a critical role in infection by protecting the parasites from oxidative stress and increasing IFN1-b expression.-

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“…It has been reported that during CHIKV infection it is antagonized by nsP2 (see above; Fros et al 2015). Interestingly, a study by Rathore and coworkers revealed that nsP4 may also have a role in this process.…”

Section: Nsp4mentioning

confidence: 93%

“…As transient expression of CHIKV glycoproteins triggers UPR, the ability to suppress UPR is associated with the nonstructural region. Indeed, it was also shown that transient expression of N-terminally tagged nsP2 (from CHIKV of WA genotype) is sufficient for UPR suppression and that mutations, known to suppress cytotoxic properties of nsP2, make the protein unable to interfere with UPR (Fros et al 2015).…”

Section: Interaction With Other Viral and Host Proteinsmentioning

confidence: 97%

Functions of Chikungunya Virus Nonstructural Proteins

Ahola

Merits

2016

Chikungunya Virus

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IntroductionThe nonstructural proteins (nsPs) of chikungunya virus (CHIKV) and other alphaviruses are encoded at the 5′ region of the positive-strand viral RNA genome, and thus translated directly after the entry of the RNA into the cytoplasm, to enable the immediate start of the RNA replication process. Many of the replicative functions remain poorly studied for CHIKV nsPs, but have been earlier characterized for proteins from other alphaviruses, primarily Semliki Forest virus (SFV) and Sindbis virus (SINV; Kaariainen and Ahola 2002). It is expected that the basic biochemical functions are conserved in CHIKV proteins, the amino acid (aa) sequence of which are on average 71 % identical with the more closely related SFV nsPs. Therefore we review data from CHIKV whenever available, but also draw heavily on other alphaviruses, and point out the uncertainties regarding the functions of CHIKV proteins.Secondly, the nsPs have many functions in host-virus interactions, including the evasion of antiviral responses. Clearly, these functions are more virus-(and/or celltype) specific in nature, so care needs to be taken in making inferences regarding CHIKV. For instance, it has been shown that hnRNP K, a cellular protein identified as a pro-viral (essential) factor for CHIKV replication (Bourai et al. 2012) acts as a negative effector of replication for SFV (Varjak et al. 2013). A primary division within the mammalian/avian alphaviruses lies between the Old World viruses (e.g., CHIKV, SFV, and SINV) and the New World viruses (e.g., Venezuelan equine encephalitis virus, VEEV), and these two groups display quite different modes of T. Ahola

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Chikungunya virus non-structural protein 2-mediated host shut-off disables the unfolded protein response

Cited by 62 publications

References 53 publications

The Antiviral Alkaloid Berberine Reduces Chikungunya Virus-Induced Mitogen-Activated Protein Kinase Signaling

The Antiviral Alkaloid Berberine Reduces Chikungunya Virus-Induced Mitogen-Activated Protein Kinase Signaling

The integrated endoplasmic reticulum stress response in Leishmania amazonensis macrophage infection: the role of X‐box binding protein 1 transcription factor

Functions of Chikungunya Virus Nonstructural Proteins

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