A speed-fidelity trade-off determines the mutation rate and virulence of an RNA virus

William Fitzsimmons; Robert J. Woods; John T. McCrone; Andrew Woodman; Jamie J. Arnold; Madhumita Yennawar; Richard Evans; Craig E. Cameron; Adam S. Lauring

The 3Dpol cDNA was prepared from purified viral RNA by RT-PCR and sequenced. The I331F 460 and P356S substitutions were identified together in one experiment, and the P356S substitution 461 was identified in a second (see Table S4 ). The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 19 were verified by DNA sequencing. PV 3Dpol RdRps were expressed and purified as described 468 previously [65, 66] . The sym-sub assays used to measure assembly/elongation kinetics of 469 purified RdRp on a defined template were performed as described in [6, 30, 67] . All assays had 1 470 µM primer/template and 2 µM enzyme. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/309880 doi: bioRxiv preprint 20 bilateral hindlimb paralysis. Over 90% of all assessments were performed by members of the 494 University veterinary technical staff, who were blinded to the hypotheses and expected 495 outcomes of the studies. All surviving animals were euthanized after 12 days. These endpoints 496 were also used to calculate the PD50 using the Spearman-Karber method (Table S3) .