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. Author manuscript; available in PMC: 2022 Sep 3.
Published in final edited form as: Science. 2021 Sep 1;373(6559):1161–1166. doi: 10.1126/science.abb3356

Fig. 3. tRNAGly sequestration by CMT-mutant GlyRS induces ribosome stalling.

Fig. 3.

(A) Size-exclusion chromatography of purified recombinant human GlyRS proteins. D:M= dimer:monomer ratio. (B) Kon and Koff values of tRNAGly-GCC binding and release to dimer and monomer forms of the indicated GlyRS variants. The (percentage) denotes the frequency of a measured value. (C) Quantification of tRNAGly bound to GlyRS in tRNAGly:GlyRS complexes immunoprecipitated from whole brains of GarsC201R/+ and WT littermate control mice. tRNAGly/GlyRS ratio of WT is set as 100%; n=5 independent experiments; *p<0.05 by one-sample t-test. (D) Hanging time in the inverted grid test of male Gtpbp2+/? or −/−; Gars+/+ (control), Gtpbp2+/?; GarsC201R/+, and Gtpbp2−/−; GarsC201R/+ littermate mice at 4, 5, 6, 7 and 8 weeks of age. n=15–28 mice per genotype group; ***p<0.0005 by one-sample t-test and two-tailed unpaired t-test with Bonferroni correction per time point. (E) Nerve conduction velocity of the sciatic nerve at 8 weeks of age. n=13–20 mice per genotype group; ***p<0.0001 by Brown-Forsythe and Welch ANOVA. (F) Axon number in the motor branch of the femoral nerve at 8 weeks of age. n=8–13 per genotype group; ***p<0.0001 by one-way ANOVA with Tukey’s multiple comparisons test. Graphs represent mean ± SEM.