We have identified four genes (
vha-5,
vha-6,
vha-7, and
unc-32) coding for vacuolar-type proton-translocating ATPase (V-ATPase) subunit a in Caenorhabditis elegans, the first example of four distinct isoforms in eukaryotes. Their products had nine putative transmembrane regions, exhibited 43-60% identity and 62-84% similarity with the bovine subunit al isoform, and retained 11 amino acid residues essential for yeast V-ATPase activity (Leng, X. H., Manolson, M. F., and Forgac, M. (1998) J. Biol. Chem. 273, 6717-6723). The similarities, together with the results of immunoprecipitation, suggest that these isoforms are components of V-ATPase. Transgenic and immunofluorescence analyses revealed that these genes were strongly expressed in distinct cells;
vha-5 was strongly expressed in an H-shaped excretory cell,
vha-6 was strongly expressed in intestine,
vha-7 was strongly expressed in hypodermis, and
unc-32 was strongly expressed in nerve cells. Furthermore, the
vha-7 and
unc-32 genes were also expressed in the uteri of hermaphrodites. RNA interference analysis showed that the double-stranded RNA for
unc-32 caused embryonic lethality similar to that seen with other subunit genes (
vha-1,
vha-4, and
vha-11) (Oka, T., and Futai, M. (2000) J. Biol. Chem. 275, 29556-29561). The progenies of worms injected with the
vha-5 or
vha-6 double-stranded RNA became died at a specific larval stage, whereas the
vha-7 double-stranded RNA showed no effect on development. These results suggest that V-ATPases with these isoforms generate acidic compartments essential for worm development in a cell-specific manner.