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. 2022 Dec 22;51(1):315–336. doi: 10.1093/nar/gkac1177

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — nsp10 stimulates hydrolysis without affecting processivity or substrate specificity. (A) Effect of nsp10 concentration on the kinetics of nsp14-catalyzed dsRNA hydrolysis. Reactions contained fixed nsp14 (0.1 μM) and varied nsp10 (0–2 μM) concentrations. In all cases, nsp14 was pre-mixed with nsp10 on ice 5 min prior to adding to the reaction. Hydrolysis of dsRNA substrate (*P10A-P40:T57) was monitored over a 30 min time course as indicated. Product analysis is shown. (B) Effect of nsp10 concentration on the kinetics of nsp14-catalyzed hydrolysis of ssRNA. Reactions were as in panel A except a ssRNA substrate (*P10A) was used. Product analysis is shown. Products shorter than n – 4 are not observed when the 10-nt ssRNA is used. (C, D) Analysis of the kinetics of product formation for both dsRNA (panel C) and ssRNA (panel D) at varying concentrations of nsp10, error bars indicate mean ± SD (n = 3).