The viral capping enzyme nsP1: a novel target for the inhibition of chikungunya virus infection

Delang, Leen; Li, C.; Taş, Ali; Quérat, Gilles; Albulescu, Irina C.; Burghgraeve, Tine De; Segura, Nidya Alexandra; Gigante, Alba; Piorkowski, Géraldine; Decroly, Étienne; Jochmans, Dirk; Canard, Bruno; Snijder, Eric J.; Pérez‐Pérez, María‐Jesús; Hemert, Martijn J. van; Coutard, Bruno; Leyssen, Pieter; Neyts, Johan

doi:10.1038/srep31819

Cited by 95 publications

(111 citation statements)

References 34 publications

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“…Lately, there has been a surge in the development of novel assays for nsP1 enzymatic activity to accentuate the pursuit of novel antiviral compounds. Previous efforts for the development of the assays for the biochemical characterization and inhibition assays against nsP1 were focussed on the GT step of the nsP1 enzymatic reaction . Gel‐based approach for inhibition study used a fluorescently labeled GTP (GTP‐ATTO) for detection or western blot/ELISA uses anti‐m 3 G/m 7 G‐cap monoclonal antibodies for quantification of nsP1 enzymatic activity .…”

mentioning

confidence: 99%

Inhibition of Chikungunya virus by an adenosine analog targeting the SAM‐dependent nsP1 methyltransferase

2019

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Alphaviruses, including Chikungunya (CHIKV) and Venezuelan equine encephalitis virus (VEEV), are among the leading causes of recurrent epidemics all over the world. Alphaviral nonstructural protein 1 (nsP1) orchestrates the capping of nascent viral RNA via its S‐adenosyl methionine‐dependent N‐7‐methyltransferase (MTase) and guanylyltransferase activities. Here, we developed and validated a novel capillary electrophoresis (CE)‐based assay for measuring the MTase activity of purified VEEV and CHIKV nsP1. We employed the assay to assess the MTase inhibition efficiency of a few adenosine analogs and identified 5‐iodotubercidin (5‐IT) as an inhibitor of nsP1. The antiviral potency of 5‐IT was evaluated in vitro using a combination of cell‐based assays, which suggest that 5‐IT is efficacious against CHIKV in cell culture (EC50: 0.409 µm).

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confidence: 99%

Inhibition of Chikungunya virus by an adenosine analog targeting the SAM‐dependent nsP1 methyltransferase

2019

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“…The activities of CHIKV nsP2 are relatively easy to analyze using purified recombinant proteins, and very recently, an assay for inhibitors of CHIKV nsP1 was developed (21). A combination of cell-based and cell-free assays was successfully used to identify inhibitors targeting nsP1 of CHIKV (22). In addition, the activities of ns proteins can indirectly be analyzed using recently developed CHIKV trans-replication system or an assay based on the use of extracts derived from infected cells (23,24).…”

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confidence: 99%

Design and Validation of Novel Chikungunya Virus Protease Inhibitors

Das

Puusepp

Varghese

et al. 2016

Antimicrob Agents Chemother

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dChikungunya virus (CHIKV; genus Alphavirus) is the causative agent of chikungunya fever. CHIKV replication can be inhibited by some broad-spectrum antiviral compounds; in contrast, there is very little information about compounds specifically inhibiting the enzymatic activities of CHIKV replication proteins. These proteins are translated in the form of a nonstructural (ns) P1234 polyprotein precursor from the CHIKV positive-strand RNA genome. Active forms of replicase enzymes are generated using the autoproteolytic activity of nsP2. The available three-dimensional (3D) structure of nsP2 protease has made it a target for in silico drug design; however, there is thus far little evidence that the designed compounds indeed inhibit the protease activity of nsP2 and/or suppress CHIKV replication. In this study, a set of 12 compounds, predicted to interact with the active center of nsP2 protease, was designed using target-based modeling. The majority of these compounds were shown to inhibit the ability of nsP2 to process recombinant protein and synthetic peptide substrates. Furthermore, all compounds found to be active in these cell-free assays also suppressed CHIKV replication in cell culture, the 50% effective concentration (EC 50 ) of the most potent inhibitor being ϳ1.5 M. Analysis of stereoisomers of one compound revealed that inhibition of both the nsP2 protease activity and CHIKV replication depended on the conformation of the inhibitor. Combining the data obtained from different assays also indicates that some of the analyzed compounds may suppress CHIKV replication using more than one mechanism. C hikungunya virus (CHIKV; genus Alphavirus, family Togaviridae) is the causative agent of chikungunya fever, a disease that has affected millions in the last decade. It is characterized by high fever, arthralgia, myalgia, headache, and rash. The disease is usually self-limiting; however long-lasting chronic symptoms are observed in nearly 50% of CHIKV-infected patients (1). As there is no approved vaccine or specific licensed antiviral compounds (2), the treatment of CHIKV infection is largely based on relief of symptoms.CHIKV infection can be inhibited by targeting host factors essential for virus infection. Targeting of several metabolic pathways has revealed anti-CHIKV effects of several licensed drugs (3). Compounds most likely targeting virus entry or host cell-specific components required for virus infection have been described (4-8). Several nucleosides or nucleotides, acting as pseudosubstrates for CHIKV RNA-dependent RNA polymerase and/or using another, more general mechanism of action, have shown anti-CHIKV activity (9). In addition, anti-CHIKV effects have been described for several groups of novel synthetic compounds (10-12).Computer-aided design based on molecular docking and molecular dynamics simulations and pharmacophore approach allows identifying potential in silico hits as active inhibitors for different CHIKV replicase proteins. This approach, however, requires the three-dimensional (3D) structure...

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“…Due to its unique features, nsP1 is an appealing target for specific antivirals. However, the lack of structural information and the propensity for membrane association (see below) have impeded studies of the protein, and only recently have specific inhibitors been described (Delang et al 2016). …”

Section: Nsp1mentioning

confidence: 98%

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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The viral capping enzyme nsP1: a novel target for the inhibition of chikungunya virus infection

Cited by 95 publications

References 34 publications

Inhibition of Chikungunya virus by an adenosine analog targeting the SAM‐dependent nsP1 methyltransferase

Inhibition of Chikungunya virus by an adenosine analog targeting the SAM‐dependent nsP1 methyltransferase

Design and Validation of Novel Chikungunya Virus Protease Inhibitors

Functions of Chikungunya Virus Nonstructural Proteins

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