Observing inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution.
Nucleic Acids Res
; 51(17): 9266-9278, 2023 09 22.
Article
em En
| MEDLINE
| ID: mdl-37560916
ABSTRACT
The genome of SARS-CoV-2 encodes for a helicase (nsp13) that is essential for viral replication and highly conserved across related viruses, making it an attractive antiviral target. Here we use nanopore tweezers, a high-resolution single-molecule technique, to gain detailed insight into how nsp13 turns ATP-hydrolysis into directed motion along nucleic acid strands. We measured nsp13 both as it translocates along single-stranded DNA or unwinds double-stranded DNA. Our data reveal nsp13's single-nucleotide steps, translocating at â¼1000 nt/s or unwinding at â¼100 bp/s. Nanopore tweezers' high spatiotemporal resolution enables detailed kinetic analysis of nsp13 motion. As a proof-of-principle for inhibition studies, we observed nsp13's motion in the presence of the ATPase inhibitor ATPγS. We construct a detailed picture of inhibition in which ATPγS has multiple mechanisms of inhibition. The dominant mechanism of inhibition depends on the application of assisting force. This lays the groundwork for future single-molecule inhibition studies with viral helicases.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
SARS-CoV-2
Limite:
Humans
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article