Antiviral Research, Journal Year: 2024, Volume and Issue: 232, P. 106037 - 106037
Published: Nov. 13, 2024
Language: Английский
Journal of Biological Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 108378 - 108378
Published: March 1, 2025
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: Feb. 6, 2024
Abstract To better understand viral pathogenesis, host-virus interactions, and potential therapeutic interventions, the development of robust reverse genetics systems for SARS-CoV-2 is crucial. Here, we present a platform that enables efficient manipulation, assembly, rescue recombinant SARS-CoV-2. The versatility our system was demonstrated by generating viruses. We used this to generate N501Y Y453F spike protein mutants. Characterization studies revealed distinct phenotypic effects, impact on fitness, cell binding, replication kinetics. also investigated recently discovered priming site NSP9, which postulated produce short RNA antisense leader sequence. By introducing U76G mutation into 5’UTR, show necessary correct production genomic subgenomic RNAs, replication. In conclusion, developed provides adaptable generation viruses their comprehensive characterization. Significance statement study, versatile facilitating Demonstrating its adaptability, successfully engineered mutants, each exhibiting effects novel negative sense demonstrating role in This straightforward genetic therefore powerful tool advancing understanding biology.
Language: Английский
Citations
3
Structure, Journal Year: 2024, Volume and Issue: 32(9), P. 1301 - 1321
Published: Sept. 1, 2024
Language: Английский
Citations
3
International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(11), P. 6120 - 6120
Published: June 1, 2024
Carnivorous pitcher plants from the genus Nepenthes are renowned for their ethnobotanical uses. This research explores therapeutic potential of miranda leaf extract against nonstructural protein 9 (Nsp9) SARS-CoV-2 and in treating human non-small cell lung carcinoma (NSCLC) lines. Nsp9, essential RNA replication, was expressed purified, its interaction with ssDNA assessed. Initial tests myricetin oridonin, known targeting ssDNA-binding proteins respectively, did not inhibit activity Nsp9. Subsequent screenings various N. extracts identified those using acetone, methanol, ethanol as particularly effective disrupting Nsp9's activity, evidenced by electrophoretic mobility shift assays. Molecular docking studies highlighted stigmast-5-en-3-ol lupenone, major components miranda, inhibitors. The cytotoxic properties were examined across NSCLC lines H1975, A549, H838, focusing on survival, apoptosis, migration. Results showed a dose-dependent effect following order: H1975 > A549 H838 cells, indicating specificity. Enhanced anticancer effects observed when combined afatinib, suggesting synergistic interactions. Flow cytometry indicated that could induce G2 cycle arrest potentially inhibiting cancer proliferation. Gas chromatography-mass spectrometry (GC-MS) enabled tentative identification 19 most abundant compounds miranda. These outcomes underscore dual utility managing infection through Nsp9 inhibition offering benefits carcinoma. results significantly broaden medical applications extract, use only traditional remedies but also prospective treatment pulmonary diseases. Overall, our findings position promising source natural therapeutics antiviral therapies, warranting further investigation into molecular mechanisms clinical applications.
Language: Английский
Citations
1
Molecular Therapy — Nucleic Acids, Journal Year: 2024, Volume and Issue: 35(3), P. 102304 - 102304
Published: Aug. 15, 2024
Nanobodies are emerging as critical tools for drug design. Several have been recently created to serve inhibitors of severe acute respiratory syndrome coronavirus s (SARS-CoV-2) entry in the host cell by targeting surface-exposed spike protein. Here we established a pipeline that instead targets highly conserved viral proteins made only after into when SARS-CoV-2 RNA-based genome is translated. As proof principle, designed nanobodies against non-structural protein (Nsp)9, which required replication. One these anti-Nsp9 nanobodies, 2NSP23, previously characterized using immunoassays and nuclear magnetic resonance spectroscopy epitope mapping, was expressed found block replication specifically. We next encapsulated 2NSP23 nanobody lipid nanoparticles (LNPs) mRNA. show this nanobody, hereby referred LNP-mRNA-2NSP23, internalized translated cells suppresses multiple variants, seen qPCR RNA deep sequencing. These results corroborated three-dimensional reconstituted human epithelium kept at air-liquid interface mimic outer surface lung tissue. observations indicate LNP-mRNA-2NSP23 and, translation, it inhibits Nsp9 living cells. speculate may be an innovative strategy generate novel antiviral drugs efficient across coronaviruses.
Language: Английский
Citations
1
Molecular Cell, Journal Year: 2023, Volume and Issue: 83(21), P. 3758 - 3760
Published: Nov. 1, 2023
Language: Английский
Citations
1
Proteins Structure Function and Bioinformatics, Journal Year: 2024, Volume and Issue: 92(11), P. 1308 - 1317
Published: July 3, 2024
ABSTRACT The ongoing global pandemic of the coronavirus 2019 (COVID‐19) disease is caused by virus SARS‐CoV‐2, with very few highly effective antiviral treatments currently available. machinery responsible for replication and transcription viral RNA during infection made up several important proteins. Two these are nsp12, catalytic subunit polymerase, nsp9, a cofactor nsp12 involved in capping priming RNA. While recent studies have determined structural details interaction nsp9 context capping, biochemical or biophysical In this study, we used combination surface plasmon resonance (SPR) experiments, size exclusion chromatography (SEC) assays to identify specific residues that critical binding as well RNAylation, both which essential process. Our data indicate dimerization unlikely play significant functional role virus. We confirm set recently discovered peptides inhibit nsp9–nsp12 specifically nsp9; however, find do not impact RNAylation. summary, our results implications future drug discovery efforts combat SARS‐CoV‐2 any newly emerging coronaviruses.
Language: Английский
Citations
0
The Journal of Physical Chemistry B, Journal Year: 2024, Volume and Issue: unknown
Published: Oct. 2, 2024
Several antiviral therapeutic approaches have been targeted toward the RNA-dependent RNA polymerase (RdRp) complex that is involved in viral genome replication. In SARS-CoV-2, although RdRp a multiprotein complex, focus has on ligand binding catalytic core (nonstructural protein nsp12), and not functional dynamics. this study, we conformational ensembles of their modulation by presence RNA, performing comprehensive microsecond-scale atomistic simulations apo- RNA-bound complex. We delineate differential impact constituent proteins, such as polymorphisms, dominant segment-specific fluctuations, switch dynamical crosstalk within distinguish signatures nsp7, nsp8, nsp12 apo-state are reduced appear to "prime" for activity. Importantly, identify unique structural malleability nsp8 with high heterogeneity apo state, especially at three sites (Y71 nsp8A, D52 A66 nsp8B). Our work highlights implications polymorphism structures reveals possibilities development allosteric inhibitors.
Language: Английский
Citations
0
Antiviral Research, Journal Year: 2024, Volume and Issue: 232, P. 106037 - 106037
Published: Nov. 13, 2024
Language: Английский
Citations
0