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Observing Inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution.

Sinduja K Marx, Keith J Mickolajczyk, Jonathan M Craig, Christopher A Thomas, Akira M Pfeffer, Sarah J Abell, Jessica D Carrasco, Michaela C Franzi, Jesse R Huang, Hwanhee C Kim, Henry Brinkerhoff, Tarun M Kapoor, Jens H Gundlach, and Andrew H Laszlo. "Observing Inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution." Nucleic Acids Research. August 2023.

Here we used the nanopore as a single-molecule tool to observe the motion of the SARS-CoV-2 helicase. By observing how the helicase walked along single-stranded DNA and unwound double-stranded DNA, we could determine its mechanism of motion. We also observed how it is affected by an inhibitory molecule. Single ATPase turnovers are clearly resolved even as nsp13 translocates at speeds in excess of 1000 nucleotides per second. This is nearly three orders of magnitude beyond what is possible with other single-molecule techniques.