Identification of potent inhibitors of arenavirus and SARS-CoV-2 exoribonucleases by fluorescence polarization assay

Hernández, Sergio; Feracci, Mikaël; Jesus, Carolina Trajano De; Kazzi, Priscila El; Kaci, Rafik; Garlatti, Laura; Mondielli, Clémence; Bailly, Fabrice; Cotelle, Philippe; Touret, Franck; Lamballerie, Xavier De; Coutard, Bruno; Decroly, Étienne; Canard, Bruno; Ferrón, François; Alvarez, Karine

doi:10.1016/j.antiviral.2022.105364

Cited by 3 publications

(3 citation statements)

References 47 publications

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“…It would be interesting to evaluate whether the ExoN activity of the NP from mammarenaviruses may be involved in an RNA proof-reading function as well, as recently suggested for the NP from MOPV and LCMV [54]. In sum, this work contributes to the understanding of the JUNV NP functions and highlights the relevance of the DEDDh motif of this protein, which, as previously shown for SARS-CoV-2 and OW LASV and LCMV [55][56][57], might be evaluated as potential target for the design of antiviral drugs to combat AHF, a concept that could be extended to other pathogenic NW mammarenaviruses.…”

Section: Discussionsupporting

confidence: 64%

Nuclease Activity of the Junín Virus Nucleoprotein C-Terminal Domain

Sierra,

Loureiro,

Esperante

et al. 2023

Viruses

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The mammarenavirus Junín (JUNV) is the causative agent of Argentine hemorrhagic fever, a severe disease of public health concern. The most abundant viral protein is the nucleoprotein (NP), a multifunctional, two-domain protein with the primary role as structural component of the viral nucleocapsids, used as template for viral polymerase RNA synthesis activities. Here, we report that the C-terminal domain (CTD) of the attenuated Candid#1 strain of the JUNV NP can be purified as a stable soluble form with a secondary structure in line with known NP structures from other mammarenaviruses. We show that the JUNV NP CTD interacts with the viral matrix protein Z in vitro, and that the full-length NP and Z interact with each other in cellulo, suggesting that the NP CTD is responsible for this interaction. This domain comprises an arrangement of four acidic residues and a histidine residue conserved in the active site of exoribonucleases belonging to the DEDDh family. We show that the JUNV NP CTD displays metal-ion-dependent nuclease activity against DNA and single- and double-stranded RNA, and that this activity is impaired by the mutation of a catalytic residue within the DEDDh motif. These results further support this activity, not previously observed in the JUNV NP, which could impact the mechanism of the cellular immune response modulation of this important pathogen.

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Section: Discussionsupporting

confidence: 64%

Nuclease Activity of the Junín Virus Nucleoprotein C-Terminal Domain

Sierra,

Loureiro,

Esperante

et al. 2023

Viruses

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“…We selected positive supernatants indicative of the presence of anti-SARS-CoV-2 Abs and cultured polyclonal B cells until they reached a monoclonal stage. Subsequently, we evaluated 445 clone supernatants containing naturally produced anti-SARS-CoV-2 Abs for their ability to protect VeroE6/ TMPRSS2 cells from the cytopathic effect induced by SARS-CoV-2 infection, following a standardized screening assay [ 16 , 18 ]. Each clone supernatant was individually assessed for its impact on the infectivity of the prototype D614G variant, as well as Delta and first Omicron variants (BA.1).…”

Section: Resultsmentioning

confidence: 99%

Neutralizing and enhancing monoclonal antibodies in SARS-CoV-2 convalescent patients: lessons from early variant infection and impact on shaping emerging variants

Coutant,

Touret,

et al. 2024

Emerging Microbes & Infections

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Serological studies of COVID-19 convalescent patients have identified polyclonal lineage-specific and cross-reactive antibodies (Abs), with varying effector functions against virus variants. Individual specificities of anti-SARS-CoV-2 Abs and their impact on infectivity by other variants have been little investigated to date. Here, we dissected at a monoclonal level neutralizing and enhancing Abs elicited by early variants and how they affect infectivity of emerging variants. B cells from 13 convalescent patients originally infected by D614G or Alpha variants were immortalized to isolate 445 naturally-produced anti-SARS-CoV-2 Abs. Monoclonal antibodies (mAbs) were tested for their abilities to impact the cytopathic effect of D614G, Delta, and Omicron (BA.1) variants. Ninety-eight exhibited robust neutralization against at least one of the three variant types, while 309 showed minimal or no impact on infectivity. Thirty-eight mAbs enhanced infectivity of SARS-CoV-2. Infection with D614G/Alpha variants generated variant-specific (65 neutralizing Abs, 35 enhancing Abs) and cross-reactive (18 neutralizing Abs, 3 enhancing Abs) mAbs. Interestingly, among the neutralizing mAbs with cross-reactivity restricted to two of the three variants tested, none demonstrated specific neutralization of the Delta and Omicron variants. In contrast, cross-reactive mAbs enhancing infectivity ( n = 3) were found exclusively specific to Delta and Omicron variants. Notably, two mAbs that amplified in vitro the cytopathic effect of the Delta variant also exhibited neutralization against Omicron. These findings shed light on functional diversity of cross-reactive Abs generated during SARS-CoV-2 infection and illustrate how the balance between neutralizing and enhancing Abs facilitate variant emergence.

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“…Any alteration of the remaining ion impairs greatly theses activities [2]. We present a global study aiming to characterize the assembly of the NP [3], through flexible domains [4], a step critical for vRNApackaging and the polsitioning of L for vRNA replication, as well as using a combined approach of biophysical screening, crystallography and in silico docking, identifying active compounds against both nucleases [5]. Crystal structures of the nucleases domain complexed with several compounds were obtained [67].…”

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confidence: 99%

Structural characterisation and inhibition of arenavirus replication complex elements: assembly, function and inhibition of embedded nucleases

Hernandez,

Papageorgiou,

Spiliopoulou

et al. 2023

Acta Cryst Sect A

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Identification of potent inhibitors of arenavirus and SARS-CoV-2 exoribonucleases by fluorescence polarization assay

Cited by 3 publications

References 47 publications

Nuclease Activity of the Junín Virus Nucleoprotein C-Terminal Domain

Nuclease Activity of the Junín Virus Nucleoprotein C-Terminal Domain

Neutralizing and enhancing monoclonal antibodies in SARS-CoV-2 convalescent patients: lessons from early variant infection and impact on shaping emerging variants

Structural characterisation and inhibition of arenavirus replication complex elements: assembly, function and inhibition of embedded nucleases

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