Figure 1.

Loss of function alleles of rswl and scu cause lethality in flies. (A) Schematic showing the domain structure of Roswell, Scully, and Mulder and their homology in humans. The mitochondrial targeting sequence (MTS, blue) was predicted using MitoProt server, and the domain boundaries were predicted using Clustal Omega based on human homologs [30,31]. MTase: methyltransferase. (B–D) CRISPR/Cas9 induced loss of scu caused lethality. (B) Antibodies raised against Scu indicate all five scu alleles appear protein null via Western blot compared to the wild-type (y w), including ubiquitous scu^GL RNAi expression. (C,D) scu^KO7 and scu^KO11 had delayed pupation (C) and failed to eclose (D) at room temperature. (E–G) The three rswl alleles did not have detectable protein on Western blot. Ubiquitous rswl^GD RNAi expression shows reduced protein levels (E). (F) rswl mutant larvae eventually pupate but had delayed development, and none enclosed at room temperature (G). CVA: anti-ATP synthase. s.d. was calculated using GraphPad PRISM (C,D,F,G).