Fig. 1: WAH-1 interacts with CPS-6 in mitochondria and promotes PME and α-synuclein-induced DA neuronal death.
a A CPS-6/WAH-1 structural model generated via ZDOCK, in which a CPS-6 dimer (blue) interacts with one WAH-1 monomer (brown), with the Mg2+ ion (yellow) interacting with the catalytic His-Me finger motif (green) in CPS-6. Arg473 (red) in WAH-1, shown in an enlarged view, is identified as a key interfacial residue between WAH-1 and CPS-6. b A schematic diagram of the GFP11x7 tag and GFP1–10 tag inserted into the cps-6 and wah-1 loci, respectively, by genome editing. The sequence of GFP11 is shown. In the GFP11x7::CPS-6(Δ) strain, CPS-6 residues 114–295 are deleted. c, e Representative GFP, TMRE and merged images of embryos (c) or GFP images of embryos (e) carrying the indicated knockins at the stage of 6 to 8 cells without (c) or with the indicated treatments (e). The mock treatment was M9 buffer. Scale bars, 10 µm. d, f Quantification of relative GFP fluorescence intensity of embryos from the indicated strains with or without the indicated treatment, n ≥ 25. Data are mean ± SEM. g A diagram of C. elegans mtDNA, the uaDf5 deletion, primers and expected sizes of PCR products in each round of the nested PCR assays. h–j Representative gel images of PCR products from 15 cross-fertilized embryos (MTR-positive) at the indicated stages (h, i) or 100-cell stage (j) from the indicated crosses with MTR-stained males. PCR controls are N2 or uaDf5/+ embryos. k, l Quantification of MTR-stained paternal mitochondrial clusters in 64-cell embryos from the indicated crosses with MTR-stained males without or with the indicated treatment. Data are mean ± SEM; n = 20 (k) or 15 (l) per cross. m, n Comparison of DA neuronal loss in adult day 2 baIn11 animals in the indicated genetic background without or with the indicated treatment. Data are mean ± SEM, n = 50 animals per experiment in 4 independent experiments for each genotype and condition. *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant (one-way ANOVA; d, f, k–n). wah-1(sm342[R437E]) and cps-6(tm3222) alleles were used.
