The sex-determining gene
fem-1 is one of three fem genes required for male development. Its product, FEM-1, is a 656-amino acid protein containing seven copies of the ANK motif near its N-terminus, and block of sequence resembling a repeat found in kinesin light chains near its C-terminus. A mouse homolog of
fem-1, mFem1a, when expressed behind the
fem-1 promoter, causes defects in sexually dimorphic structures of transgenic
fem-1 animals. This effect is consistent with mFem1a having weak
fem-1 (+) activity. We are attempting to improve upon this weak rescue by optimizing transgene expression, using synthetic introns and different promoters. Using the sequence conservation between the mouse and worm genes as a guide, we designed primers for degenerate PCR, to isolate
fem-1 homologs from other species. To date, we have cloned and sequenced a 300 bp fragment of the C. briggsae
fem-1 homolog. Once we have recovered a genomic clone of the complete C. briggsae
fem-1 gene, we will test whether it is capable of rescuing C. elegans
fem-1 mutants. We also plan to use RNA-interference to determine whether the gene is involved in C. briggsae sex determination. The comparison of the C. elegans, C. briggsae, and mouse sequences and rescuing activities will reveal regions of
fem-1 which are important for its activity. In addition, conservation of function implies that some of the interactions involving FEM-1 have also been conserved during evolution.