Fig. 5. gld-2 and nos-3 function to promote GLD-1 protein accumulation. (A) Diagram of NOS-3 protein drawn to scale showing the location of the lesions associated with the nos-3 alleles obtained in the genetic screen described (Fig. 1). Shaded boxes represent the two putative zinc fingers that are similar to Drosophila Nanos. The oz233, oz235, oz239 and oz240 alleles are associated with nonsense mutations predicted to result in truncated proteins 308, 177, 355 and 568 amino acids in length respectively, as compared to 871 amino acids of full-length NOS-3 (Kraemer et al., 1999). The oz231 allele is associated with a 139 base pair deletion (open box), deleting amino acids 427-473, as well as changing the reading frame, therefore adding 39 amino acids (filled box) before encountering a stop codon. All lesion locations refer to the previously published splice form of nos-3 (Kraemer et al., 1999), however we have identified two alternative splices that affect exons five and seven. The alternative splice sites have also been identified in large scale cDNA sequencing efforts and are noted (http://www.wormbase.org, release WS100, May 2003), with nos-3b corresponding to the previously identified splice form (Kraemer et al., 1999). (B-F) GLD-1 protein accumulation (red) and DAPI (blue) in dissected gonad arms of (B) wild-type, (C) gld-2(q497), (D) nos-3(oz231), (E) gld-2(q497); nos-3(oz231) and (F) gld-2(q497); nos-3(oz231); unc-32(e189) glp-1(q175) animals one day past L4 at 20°C. The distal end is to the left and the proximal portion of each arm is not shown. Wild-type (B) and mutant animals (C-F) were dissected, fixed and stained together and pictures taken with the same settings and processed identically (see Materials and methods). Scale bar: 20 µm.