Positional cloning, phylogenetic analysis, and alignment of WSC-related sequences. (A) The schematic shows a set of markers on the left arm of chromosome three. The recombination frequency between wsc and these markers is indicated at the top. Cosmids (a–h) encompassing contig 40 were assessed for their ability to complement the wsc mutant. Complementing cosmids (e and f) are drawn as solid lines and noncomplementing cosmids are drawn as dotted lines. Transformation with DNA fragments containing individual candidate genes identified NCU 07842.2 as wsc. (B) Alignment of WSC and selected homologous proteins. Regions predicted to encode membrane-spanning domains are highlighted in yellow (score > 500) and pink (score < 500). Identical (*), strongly conserved (:), and weakly conserved (.) residues are identified beneath the alignment. The arginine residue that is mutated in some individuals afflicted with MDDS is shaded with black. (C) Phylogenetic tree showing the relationship between WSC and homologous sequences. The tree was constructed using MrBayes3.1.2. The scale bar indicates 0.1 substitutions. Proteins that have been shown to reside in the peroxisome are shown in green and those known to target to mitochondria are shown in blue. WSC and close relatives in the Euascomycetes are labeled in red. AT, Arabidopsisthaliana; AF, Aspergillus fumigatus; AN, Aspergillus nidulans; AO, Aspergillus oryzae; CA, Candida albicans; CC, Coprinus cinereus; DD, Dictyostelium discoideum; DM, Drosophila melanogaster; GZ, Gibberella zea; HC, Histoplasma capsulatum; HS, Homo sapiens; KL, Kluyveromyces lactis; MG, Magnaporthe grisea; NC, Neurospora crassa; OS, Oryzae sativa; SC, Saccharomyces cerevisiae; SP, Schizosaccharomyces pombe; SS, Sclerotina sclerotinium; UM, Ustilago maydis; YL, Yarrowia lipolytica; XT, Xenopus tropicalis.