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Fig. 5. ehn-3 encodes an SGP-specific protein with paired zinc fingers.
(A) The ehn-3 locus. cDNA analyses identify three transcripts, each
predicted to encode a protein with multiple zinc fingers (black boxes). All
cDNAs are transpliced to the SL1 leader. ehn-3A encodes six zinc
fingers; ehn-3B1 and B2 encode four zinc fingers. The gene
predicted by the sequence consortium, called ZK616.10, corresponds to
ehn-3B. ehn-3(q689) is associated with an insertion (T) and a
C
T transition in exon 2 of ehn-3A. ehn-3(q766) deletes a region
including exon 2 from ehn-3A, the next intron and part of the first
common exon, including coding sequence for the first 14 amino acids of EHN-3B1
and B2. (B) Semi-quantitative PCR used to assess transcript levels in
ehn-3 mutants. (Left) ehn-3 PCR products were less abundant
when amplified from ehn-3 mutants than from wild type (lower band in
N2 is ehn-3B2); by contrast, hlh-2 PCR product was similar
in wild-type and ehn-3 mutants. (Right) Ratio of ehn-3 to
hlh-2 PCR products. (C) Alignment of EHN-3 putative zinc fingers.
ZNF1-ZNF3 and ZNF6 were predicted by Pfam; ZNF4 and ZNF5 were identified by
eye. (D-F) SGP-specific expression of EHN-3::GFP. (D) EHN-3::GFP transgene
composition: ehn-3 genomic DNA, including 429 bp of 5' flanking
sequence plus the entire EHN-3-coding region, was fused in frame to the
GFP-coding sequence. (E) Wild-type embryo expressing EHN-3::GFP. SGP (arrow)
has assembled into the gonadal primordium; PGC, asterisk. (F) EHN-3::GFP is
expressed in SGP (arrowhead) shortly after gonadal primordium is formed.
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