The comparison of cell fate specification in the vulva equivalence group between C. elegans and Pristionchus pacificus reveals several evolutionary differences. In P. pacificus, P8.p loses its competence to adopt vulval cell fate early in the L1 stage and can only adopt an epidermal fate. In mutagenesis screens of a total of appr. 100 000 gametes many egg-laying defective mutations were isolated. In 11 of these mutations the specification of P8.p was affected. In most mutant animals analyzed, a posterior vulva-like structure was formed by P8.p. The specification of P7.p was also affected: cell lineage analysis in 5 strains revealed that P7.p adopted the 1! fate in approximately 5% of animals. In such animals, the main invagination was generated by the progeny of P(6,7).p and P6.p had the 2! fate. The AC in these cases was displaced posteriorly so that it was now located above P7.p. Complementation tests revealed that all 11 mutations are allelic to one another and to the previously isolated mutation
Ppa-ped-4. Further genetic analysis indicated that
Ppa-ped-4 is linked to the visible mutation
Ppa-dpy-1 which has been shown to be closely linked to the homeotic gene
Ppa-lin-39. This result suggests, that
Ppa-ped-4 might be associated with, or might be a member of the Hox genes of P. pacificus. One candidate gene would be
mab-5. To test if
Ppa-ped-4 corresponds to P. pacificus
mab-5, we cloned the
Ppa-mab-5 homeodomain by PCR and isolated genomic and cDNA clones. We analyzed the complete
Ppa-mab-5 coding region in 5 of the 12
ped-4 mutant strains and found mutations in all of them indicating that
Ppa-ped-4 is identical to the homeotic gene
mab-5. To simplify nomenclature we renamed
Ppa-ped-4 as
Ppa-mab-5. When we ablated Z(1,4) in
Ppa-mab-5 mutant animals, P7.p and P8.p could differentiate into vulval cells, whereas P(5,6).p remained epidermal. These results suggest that P(7,8).p can differentiate in a gonad-independent way in
Ppa-mab-5 mutant animals, indicating that P(7,8).p are distinct from P(5,6).p. The gonad-independent differentiation properties of P(7,8).p might occur in response to an unknown signal or because of an intrinsic property of these cells to undergo vulva-like differentiation. In wild-type animals,
Ppa-mab-5 functions as part of a negative signaling system that antagonizes the differentiation properties of P(7,8).p. The
Ppa-mab-5-mediated process prevents the inappropriate differentiation of P8.p by making this cell incompetent to adopt vulval fate. This effect of
Ppa-mab-5 is eventually restricted to P8.p, as the activity in P7.p is overruled by gonadal signaling that induces vulva formation. Comparing the
mab-5 lof phenotypes, the strongest difference between species is that in
Cel-mab-5 no ectopic vulva differentiation occurs. Furthermore,
Cel-mab-5 mutant animals have VPCs that are identical in their developmental potential. In contrast, in
Ppa-mab-5 mutations P(7,8).p still differ from P(5,6).p resulting in gonad independent differentiation. This observation indicates that factors other than
mab-5 distinguish P(7,8).p from P(5,6).p or vice versa. One major function of Ppa-MAB-5 is to antagonize gonad-independent differentiation of P(7,8).p. Mechanistically, MAB-5 might act in different genetic circuits with different target genes in both species. For example,
Ppa-mab-5 might be part of a locally restricted negative signaling system that is only present in P. pacificus P(7,8).p. However, we cannot rule out that only quantitative differences of MAB-5 activity in the regulation of identical downstream target genes led to the observed competence patterns in both species.