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J. Biol. Chem., Vol. 276, Issue 35, 33079-33085, August 31, 2001
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From the Division of Biological Sciences, Institute of Scientific
and Industrial Research, Osaka University, and Core Research for
Evolutional Science and Technology (CREST) of the Japan Science and
Technology Corporation, Osaka 567-0047, Japan
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 a1 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.
The nucleotide sequences reported in this paper have been
submitted to the DDBJ/GenBankTM/EBI Data Bank
with accession numbers AB055110, AB055111, and
AB055112.
Four Subunit a Isoforms of
Caenorhabditis elegans Vacuolar H+-ATPase
CELL-SPECIFIC EXPRESSION DURING DEVELOPMENT*
*
This work was supported in part by a grant from the Ministry
of Education, Culture, Sports, Science and Technology of Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 81-6-6879-8480;
Fax: 81-6-6875-5724; E-mail: m-futai@sanken.osaka-u.ac.jp.
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