Cypovirus capsid protein VP5 has nucleoside triphosphatase activity

Yang, Jie; Qi, Qian; Li, Teng-Feng; Yang, Xueli; Won, Sok Jin; Zhou, Xi

doi:10.1007/s12250-017-3999-2

Cited by 6 publications

(9 citation statements)

References 12 publications

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“…NTPase activities were performed by measuring the released inorganic phosphate during NTP hydrolysis via a direct colorimetric assay as previously described (Yang et al 2017). The concentrations of inorganic phosphate were determined by matching the absorbance at wavelength 620 nm (A620) in a known standard inorganic phosphate curve.…”

Section: Ntpase Assaymentioning

confidence: 99%

SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts

et al. 2020

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The ongoing outbreak of Coronavirus Disease 2019 has become a global public health emergency. SARScoronavirus-2 (SARS-CoV-2), the causative pathogen of COVID-19, is a positive-sense single-stranded RNA virus belonging to the family Coronaviridae. For RNA viruses, virus-encoded RNA helicases have long been recognized to play pivotal roles during viral life cycles by facilitating the correct folding and replication of viral RNAs. Here, our studies show that SARS-CoV-2-encoded nonstructural protein 13 (nsp13) possesses the nucleoside triphosphate hydrolase (NTPase) and RNA helicase activities that can hydrolyze all types of NTPs and unwind RNA helices dependently of the presence of NTP, and further characterize the biochemical characteristics of these two enzymatic activities associated with SARS-CoV-2 nsp13. Moreover, we found that some bismuth salts could effectively inhibit both the NTPase and RNA helicase activities of SARS-CoV-2 nsp13 in a dose-dependent manner. Thus, our findings demonstrate the NTPase and helicase activities of SARS-CoV-2 nsp13, which may play an important role in SARS-CoV-2 replication and serve as a target for antivirals.

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Section: Ntpase Assaymentioning

confidence: 99%

SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts

et al. 2020

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“…Because a number of viral RNA remodelling proteins also have NTPase activity (22,34,47), we next examined if EBOV VP35 has NTPase activity that can hydrolyze NTPs (ATP, GTP, CTP and UTP) by using a sensitive colorimetric assay that measures the total amount of free orthophosphate released after ATP hydrolysis. Our data showed that MBP–VP35 hydrolyzed all four types of NTPs, with a preference on ATP, GTP and UTP over CTP (Figure 1A).…”

Section: Resultsmentioning

confidence: 99%

“…Factually, Ectropis obliqua picorna-like virus (EoV) 2C protein contains the conserved motif A, which is commonly recognized as the core NTP binding and NTPase active site of SF3 helicases (21), but the NTPase activity of EoV 2C is not dependent on this motif. In addition, the capsid protein VP5 of Helicoverpa armigera cypovirus-5 (HaCPV-5; genus Cypovirus , family Reoviridae ) has the NTPase activity, which is also Mg 2+ -dependent and can hydrolyze all kinds of NTPs and dNTPs, but contains no conserved NTPase/helicase motifs (27,47). Thus, our current study, together with the previous studies, imply that some of the NTPase and RNA remodeling activities may not strictly rely on conserved motifs in linear sequences of amino acids, but probably determined by more sophisticated active sites formed in tertiary protein structures.…”

Section: Discussionmentioning

confidence: 99%

Ebola virus VP35 has novel NTPase and helicase-like activities

Shu

Gan

Bai

et al. 2019

Nucleic Acids Research

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Ebola virus (EBOV) is a non-segmented, negative-sense RNA virus (NNSV) in the family Filoviridae , and is recognized as one of the most lethal pathogens in the planet. For RNA viruses, cellular or virus-encoded RNA helicases play pivotal roles in viral life cycles by remodelling viral RNA structures and/or unwinding viral dsRNA produced during replication. However, no helicase or helicase-like activity has ever been found to associate with any NNSV-encoded proteins, and it is unknown whether the replication of NNSVs requires the participation of any viral or cellular helicase. Here, we show that despite of containing no conserved NTPase/helicase motifs, EBOV VP35 possesses the NTPase and helicase-like activities that can hydrolyse all types of NTPs and unwind RNA helices in an NTP-dependent manner, respectively. Moreover, guanidine hydrochloride, an FDA-approved compound and inhibitor of certain viral helicases, inhibited the NTPase and helicase-like activities of VP35 as well as the replication/transcription of an EBOV minigenome replicon in cells, highlighting the importance of VP35 helicase-like activity during EBOV life cycle. Together, our findings provide the first demonstration of the NTPase/helicase-like activity encoded by EBOV, and would foster our understanding of EBOV and NNSVs.

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“…The point mutations were introduced into the NS2 coding region by PCR-mediated mutagenesis with the appropriate primers. For expression of recombinant NS2 in sf9 cells, the ORFs of NS2 and its mutant were constructed into the vector pFastBac HTB-MBP as previously described (Yang et al 2017). The resulting plasmids were subjected to the Bac-to-Bac baculovirus expression system to express the fusion proteins with an MBP tag at the N-terminus.…”

Section: Plasmids and Rnasmentioning

confidence: 99%

“…The expression and purification of MBP alone and MBPfusion proteins were performed in sf9 cells using baculovirus expression system as previously described (Yang et al 2017). Briefly, cells were infected with MBP-tagged HCV NS2 expressing baculoviruses for 72 h. Infected cells were resuspended, lysed via sonication and then centrifuged at 11,000 9g for 30 min to remove debris.…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

Hepatitis C Virus NS2 Protein Suppresses RNA Interference in Cells

Zhou

Shu

et al. 2019

Virol. Sin.

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RNAi interference (RNAi) is an evolutionarily conserved post-transcriptional gene silencing mechanism and has been well recognized as an important antiviral immunity in eukaryotes. Numerous viruses have been shown to encode viral suppressors of RNAi (VSRs) to antagonize antiviral RNAi. Hepatitis C virus (HCV) is a medically important human pathogen that causes acute and chronic hepatitis. In this study, we screened all the nonstructural proteins of HCV and found that HCV NS2 could suppress RNAi induced either by small hairpin RNAs (shRNAs) or small interfering RNAs (siRNAs) in mammalian cells. Moreover, we demonstrated that NS2 could suppress RNAi via its direct interaction with doublestranded RNAs (dsRNAs) and siRNAs, and further identified that the cysteine 184 of NS2 is required for the RNAi suppression activity through a serial of point mutation analyses. Together, our findings uncovered that HCV NS2 can act as a VSR in vitro, thereby providing novel insights into the life cycle and virus-host interactions of HCV.Keywords Hepatitis C virus (HCV) Á NS2 Á Antiviral RNAi Á Viral suppressor of RNAi (VSR)

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Cypovirus capsid protein VP5 has nucleoside triphosphatase activity

Cited by 6 publications

References 12 publications

SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts

SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts

Ebola virus VP35 has novel NTPase and helicase-like activities

Hepatitis C Virus NS2 Protein Suppresses RNA Interference in Cells

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