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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.
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