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. 2022 Jan 14;298(3):101590. doi: 10.1016/j.jbc.2022.101590

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Mettl5 knockout mice have developmental and metabolic defects.A, levels of m⁶A (left) and m^6,6A (right), normalized to G, obtained by LC-MS/MS of 40-nt probe-purified segments of 18S rRNA surrounding the m⁶A 1832 site from the brains and livers of Mettl5^+/+, Mettl5^+/−, and Mettl5^−/− mice. n = 4, Mettl5^+/+, n = 3, Mettl5^+/−, and n = 4, Mettl5^−/−. Analyzed by one-way ANOVA comparison with Sidak test for multiple comparisons. B, pie chart of the predicted (left) and actual (right) mice born of wild type (+/+, dark blue), heterozygous (+/−, grey), and knockout (−/−, red) genotypes from heterozygous x heterozygous breeding pairs (total = 106 mice; 30 +/+, 56 +/−, 20 −/− mice). C, photograph of male littermate Mettl5^+/+, Mettl5^+/−, and Mettl5^−/− mice at 8 weeks of age. D, body weights in grams of male (top) and female (bottom) Mettl5^+/− and Mettl5^−/− mice from 4 weeks (weaning) to 10 weeks of age. n = 6 mice for male +/−, n = 4 for male −/−, n = 5 for female +/− and −/−; unpaired t test was used to compare each time point. E, volcano plots from mouse liver (left) and brain (right) RNA-seq data displaying the negative log₁₀ of the adjusted p value versus the log₂ fold change comparing the transcriptomes of Mettl5^−/− and Mettl5^+/− mice (n = 3 pairs). F, dysregulated pathways based on analysis of RNA-seq data from Mettl5^−/− and Mettl5^+/− mouse livers by the software DAVID (65). G, effects of METTL5 knockout on translation and transcription as displayed by log₂(fold change) of translation efficiency (TE) versus log₂(fold change) of reads per kilobase of transcript per million mapped reads (RPKM) from ribosome profiling input and ribosome-protected fragment sequencing of mouse livers. Case = Mettl5^−/−, Control = Mettl5^+/+. H, latency in seconds for Mettl5^+/− and Mettl5^−/− mice to fall off the rotarod in a rotarod performance test. I, total distance in centimeters travelled by Mettl5^+/− and Mettl5^−/− mice when placed in a new environment in an open field test. Unpaired t test shown. H and I, n = 11 pairs. ns: not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.005.