Fig. 1

seahorse encodes Lrrc6l. (A-A″) sea encodes a 440 amino acid protein (Ai); the N terminus contains four LRR motifs terminated by an LRR cap motif (green; Aii). The fourth LRR motif is disrupted by the sea^tg238a L93P missense mutation (pink). sea^fa20r Q201X nonsense mutation truncates the protein shortly after the coiled-coil (pink). The protein is predicted to contain three weak nuclear export signals (NES) and one strong nuclear localization signal (Aiii). (A′) Lrrc6l is conserved among species. An ortholog from T. brucei is 39% identical to amino acids 1-363 of Sea. Orthologs from Xenopus and mouse are 55% identical and 54% to amino acids 1-440 of Lrrc6l, respectively, and extend roughly 30 amino acids beyond the Lrrc6l C terminus. (A″) Alignment of the four Lrrc6l LRR motifs; the consensus Lrrc6l LRR sequence is [L/C]xxLxxLxLxxNxIxxIxxVxx(x) and is most similar to repeats in the SDS22-like subfamily (Kobe and Kajava, 2001). The core LRR consensus sequence shared among all LRR proteins, LxxLxLxxN/CxL (x can be any amino acid and L positions can be V, I or F), is underlined (Kobe and Kajava, 2001). (B-B″′) sea^fa20r is a 702C→T transition in exon 5 encoding a Q201X truncation (B,B′). sea^tg238a is a 273T→C transition in exon 3 encoding an L93P missense (B″,B″′). (C-C″′) Genotyping (C,C″) sea^fa20r (red) creates an AluI restriction site (black nucleotides), cleaving the 114 base pair (bp) PCR product into 81 and 33 bp fragments. (C′,C″′) sea^tg238a (red) abolishes an FspBI restriction site (black nucleotides), preventing cleavage of the 176 bp fragment into 114 and 62 bp fragments. (D,D′) Genotyping results. The left-most lanes contain DNA size markers with upper band at 200 bp and lower band at 100 bp.