Fig. 1. Molecular cloning of the ooc-5 locus. (A) Schematic diagram of the genetic and physical maps in the ooc-5 region. The top line shows the rol-6 unc-4 interval of the genetic map of chromosome II (LGII). Below it are shown deficiencies, with the region deleted indicated by hatched bars and the uncertainty in breakpoint shown by broken bars. The physical map of overlapping cosmids (black lines) is shown from left to right: F07H5, T14D7, T07D4, B0457, T21B10, C18E9, F44G4, F37B12, T24B8, C07H4, T08E2. (B) Transformation rescue of ooc-5 mutants. The name of the cosmid or subfragment tested (construct) for rescue is shown on the left, with a plus indicating rescue and the number of lines rescued/total tested shown in parentheses. The extent of each construct is shown on the right. (C) Predicted open reading frames (ORFs) and cDNAs corresponding to the two smallest rescuing fragments. The predicted exons (white boxes) and introns for F44G4.1 (predicted by the C. elegans Genome Project) are shown, with the corresponding regions of identity with cDNAs indicated below. The available sequence tags of yk643a1 and yk452e10 (gray boxes; Wormbase) were used to align these cDNAs, while the SL1 5' clone, a 3' clone and yk504f1 (cross-hatched boxes) were sequenced in their entirety. The 3' cDNA clone includes 168 nucleotides of predicted intron 4. The yk504f1 sequence has 172 nucleotides of predicted intron 4 and a stretch of A residues; this is followed by sequences with identity to T23E1.2, an unrelated ORF, which suggests it is a hybrid cDNA. (D) Schematic representation of OOC-5 domains and homology with other Torsin family members. Black boxes represent predicted signal peptides and gray boxes are additional hyrdrophobic regions. The hatched regions indicate the AAA+ ATPase domain, with the position of the Walker A (A) and Walker B (B) motifs labeled. The larger isoform of OOC-5 is shown; the smaller isoform is missing amino acids 27-32.The amino acid changes associated with the three ooc-5 mutations are indicated by arrows. Percent identity/similarity between OOC-5 and other torsin family members is indicated to the right of each protein; the regions of similarity begin after the hydrophobic domain (amino acid 49 of OOC-5) and continue for the length of the entire proteins. The GenBank Accession Numbers for these sequences are: OOC-5, Z70034 (Accession Number is for cosmid C18E9; the large OOC-5 isoform is C18E9.11b, the short isoform is C18E9.11a); Torsin A, NM_000113; Torsin B, AF317129; Y37A1B.12, CAA19495; and Y37A1B.13, CAA19484.