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. 2022 Mar 31;50(7):4012–4028. doi: 10.1093/nar/gkac184

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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 7. — SARS2 interacts with METTL8-Iso1 directly but does not determine the m³C32 modification activity of METTL8-Iso1. (A) Genes encoding METTL8-Iso1-FLAG and SARS2-Myc were coexpressed in HEK293T cells. SARS2-Myc was precipitated by METTL8-Iso1-FLAG using anti-FLAG antibodies. The addition of RNase had no effect on the METTL8-Iso1 and SARS2 interaction. (B) Time-course curves of the m³C32 modification of i⁶A-hmtRNA^Ser(UCN) by METTL8-Iso1 and different concentrations of SARS2 as indicated (METTL8-Iso1 to SARS2 mole ratio 1:1, 1:2, or 1:5). Two negative controls (modification by METTL8-Iso1 alone without (METTL8-Iso1 + no tRNA) or with (METTL8-Iso1 + hmtRNA^Ser(UCN)) tRNA transcript added were included. (C) Time-course curves of the m³C32 modification of hmtRNA^Ser(UCN) transcript by METTL8-Iso1 and SARS2 (1:2). Negative controls (METTL8-Iso1 + no tRNA) represent assays without tRNA added. Modification of i⁶A-hmtRNA^Ser(UCN) by METTL8-Iso1:SARS2 (1:2) was included as a positive control. Data represent averages of two (B) or three (C) independent experiments and the corresponding SEM.