Single-molecule data on SARS-CoV-2 RTC nsp13-helicase interaction

This dataset contains high-throughput magnetic tweezers data on the polymerase-helicase coupling of the coronavirus during the nucleotide addition cycle by the polymerase (RTC) on a ss- or dsRNA template and it is linked to the publication America, Bera et al. (2026) - RNA virus polymerase-helicase coupling enables rapid elongation through duplex RNA. We collected single-molecule traces on the RNA synthesis dynamics on dsRNA by the coronavirus RTC for increasing concentrations of nsp13-helicase, varying RNA tension for only core RTC and at saturating concentration of nsp13-helicase, typically ~300 traces per condition. We also performed control measurements with poliovirus RNA polymerase, ATPase-dead nsp13, nsp13M4 and a ssRNA template. Then we performed coarse-grained dwell-time analysis on the single-molecule traces on a 10 nucleotide window, giving 4000-8000 dwell-times condition, followed by fits on these distributions for different conditions for the coronavirus polymerase (core RTC) with the helicase (nsp13). Based on the trends in the parameters of the dwell-time fit-function, we tested different models for the assembly of the RTC-nsp13 complexes connected to the RNA synthesis dynamics and obtained a mechanochemical scheme describing the mechanism behind the speed up of the nucleotide addition cycle of the RTC by nsp13-helicase.