Fig. 1.

Analysis of mutations in yeast model. Analysis of cif permutations in wYak, wMel, and wPip. Amino acid differences between homologs are highlighted with red dots for (A) Cid^wMel versus Cid^wYak and (B) Cin^wPip versus Cin^wYak. CidB^wYak contains two mutations within the DUB domain and a large inversion. CinB^wYak contains a tandem duplication yielding a frameshift and premature stop codon but no mutations in the nuclease (Nuc) domains (C). Serial dilutions of yeast-expressing constructs with analog mutations for cid (C) and cin (D). For cid, we express cidB^wPip as in previous studies (19), since it produces a strong CI-like phenotype. Introducing (V-L) and (NTΔ) individually reduces yeast toxicity relative to wild-type control, cidB^wPip. cidB^wPip(C-A) is a catalytic inactive negative control. CEN-vector is an empty pRS416gal1 negative control. For cin, (NTΔ) beginning after the tandem duplication and the wild-type sequence containing the tandem duplication reduces yeast death relative to wild-type control, cinB^wPip. (NTΔ) and (NTΔ)^CO contain native sequence or codon optimized sequence, respectively. 2μ-vector is an empty pYes2 negative control.