Coronavirus genomic nsp14‐ExoN, structure, role, mechanism, and potential application as a drug target

Tahir, Mohammed

doi:10.1002/jmv.27009

Cited by 54 publications

(56 citation statements)

References 52 publications

(127 reference statements)

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“…Recent structural analysis highlights a network of the interdomain contacts between NSP14, NSP10 and NSP12 in the context of a capping and proofreading complex ( 11 , 12 ), yet the molecular mechanisms that define these interactions are still under investigation. NSP14 is a bifunctional enzyme characterized by an N-terminal exoribonuclease domain named for the catalytic residues in the superfamily (DEDD) and a C-terminal methyltransferase (MTase) domain which catalyzes N7- guanosine methylation ( Figure 1 A) ( 13 , 14 ). NSP10 is a 139 amino acid protein that is an activator of both NSP14 and NSP16, a 2'-O-methyltransferase, which forms a crucial step in capping nascent mRNA ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

Activation of the SARS-CoV-2 NSP14 3′–5′ exoribonuclease by NSP10 and response to antiviral inhibitors

Riccio

Sullivan

Copeland

2022

Journal of Biological Chemistry

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Understanding the core replication complex of SARS-CoV-2 is essential to the development of novel coronavirus-specific antiviral therapeutics. Among the proteins required for faithful replication of the SARS-CoV-2 genome are NSP14, a bifunctional enzyme with an N-terminal 3'-to-5' exoribonuclease (ExoN) and a C-terminal N7-methyltransferase (N7-MTase), and its accessory protein, NSP10. The difficulty in producing pure, high quantities of the NSP10/14 complex has hampered the biochemical and structural study of these important proteins. We developed a straightforward protocol for the expression and purification of both NSP10 and NSP14 from E. coli and for the in vitro assembly and purification of a stoichiometric NSP10/14 complex with high yields. Using these methods, we observe NSP10 provides a 260-fold increase in k cat / K m in the exoribonucleolytic activity of NSP14 and enhances protein stability. We also probed the effect of two small molecules on NSP10/14 activity, remdesivir monophosphate and the methyltransferase inhibitor S-adenosyl homocysteine (SAH). Our analysis highlights two important factors for drug development: first, unlike other exonucleases, the monophosphate nucleoside analogue intermediate of remdesivir does not inhibit NSP14 activity; and second, SAH modestly activates NSP14 exonuclease activity. In total, our analysis provides insights for future structure-function studies of SARS-CoV-2 replication fidelity for the treatment of COVID-19.

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Section: Introductionmentioning

confidence: 99%

Activation of the SARS-CoV-2 NSP14 3′–5′ exoribonuclease by NSP10 and response to antiviral inhibitors

Riccio

Sullivan

Copeland

2022

Journal of Biological Chemistry

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“…The nsp14 exonuclease requires divalent cations like Mg 2+ , Mn 2+ , Ca 2+ , Ni 2+ , Cu 2+ , Co 2+ , or Zn 2+ for inducing structural changes and reaction activity. 16 , 17 The proofreading activity of nsp14 was associated with nsp10, and this activity could be hindered by sofosbuvir. 18 …”

Section: Introductionmentioning

confidence: 99%

Exploring the therapeutic nature of limonoids and triterpenoids against SARS-CoV-2 by targeting nsp13, nsp14, and nsp15 through molecular docking and dynamics simulations

Vardhan

Sahoo

2022

Journal of Traditional and Complementary Medicine

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Background and aim The ongoing global pandemic due to SARS-CoV-2 caused a medical emergency. Since December 2019, the COVID-19 disease is spread across the globe through physical contact and respiratory droplets. Coronavirus caused a severe effect on the human immune system where some of the non-structural proteins (nsp) are involved in virus-mediated immune response and pathogenesis. To suppress the viral RNA replication mechanism and immune-mediated responses, we aimed to identify limonoids and triterpenoids as antagonists by targeting helicases (nsp13), exonuclease (nsp14), and endoribonuclease (nsp15) of SARS-CoV-2 as therapeutic proteins. Experimental procedure In silico molecular docking and drug-likeness of a library of 369 phytochemicals from limonoids and triterpenoids were performed to screen the potential hits that binds effectively at the active site of the proteins target. In addition, the molecular dynamics simulations of the proteins and their complexes with the potential hits were performed for 100 ns by using GROMACS. Results and conclusion The potential compounds 26-deoxyactein and 25-O-anhydrocimigenol 3-O-beta- d -xylopyranoside posing strong interactions with a minimum binding energy of −10.1 and −9.5 kcal/mol, respectively and sustained close contact with nsp13 for 100 ns. The nsp14 replication fork activity was hindered by the tomentosolic acid, timosaponin A-I, and shizukaol A with the binding affinity score of −9.2, −9.2, and −9.0 kcal/mol, respectively. The nsp15 endoribonuclease catalytic residues were inhibited potentially by limonin, 25-O-anhydrocimigenol 3-O-alpha- l -arabinopyranoside, and asperagenin posing strong binding affinity scores of −9.0, −8.8, and −8.7 kcal/mol, respectively. Computationally predicted potential phytochemicals for SARS-CoV-2 are known to possess various medicinal properties.

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“… 264 , 269 , 270 When ExoN forms the ExoN-Nsp10 complex, Nsp10 does not undergo a significant conformational change, 270 and the complex structure resembles the Asp-Glu-Asp-Asp (DEDD)-type exonuclease. 264 , 270 Both possess typical DED/Edh motifs (D90/E92/E191/H268/D273) 271 and adopt similar topological folds (a central twisted β-sheet flanked by α-helices on either side). However, distinct from the DEDD-type exonuclease, Nsp14 contains two zinc-binding sites (ZnF1 and ZnF2) that are located on both sides of the β-sheets and coordinated by C207/C210/C226/H229 and H257/C261/H264/C279, respectively.…”

Section: Nonstructural Proteins Of Sras-cov-2mentioning

confidence: 99%

Structural biology of SARS-CoV-2: open the door for novel therapies

Zheng

Zeng

et al. 2022

Sig Transduct Target Ther

164

125

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Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is the causative agent of the pandemic disease COVID-19, which is so far without efficacious treatment. The discovery of therapy reagents for treating COVID-19 are urgently needed, and the structures of the potential drug-target proteins in the viral life cycle are particularly important. SARS-CoV-2, a member of the Orthocoronavirinae subfamily containing the largest RNA genome, encodes 29 proteins including nonstructural, structural and accessory proteins which are involved in viral adsorption, entry and uncoating, nucleic acid replication and transcription, assembly and release, etc. These proteins individually act as a partner of the replication machinery or involved in forming the complexes with host cellular factors to participate in the essential physiological activities. This review summarizes the representative structures and typically potential therapy agents that target SARS-CoV-2 or some critical proteins for viral pathogenesis, providing insights into the mechanisms underlying viral infection, prevention of infection, and treatment. Indeed, these studies open the door for COVID therapies, leading to ways to prevent and treat COVID-19, especially, treatment of the disease caused by the viral variants are imperative.

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Coronavirus genomic nsp14‐ExoN, structure, role, mechanism, and potential application as a drug target

Cited by 54 publications

References 52 publications

Activation of the SARS-CoV-2 NSP14 3′–5′ exoribonuclease by NSP10 and response to antiviral inhibitors

Activation of the SARS-CoV-2 NSP14 3′–5′ exoribonuclease by NSP10 and response to antiviral inhibitors

Exploring the therapeutic nature of limonoids and triterpenoids against SARS-CoV-2 by targeting nsp13, nsp14, and nsp15 through molecular docking and dynamics simulations

Structural biology of SARS-CoV-2: open the door for novel therapies

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