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Molecular Cloning of Vacuolar H+-Pyrophosphatase and
Its Developmental Expression in Growing Hypocotyl of Mung
Bean1
Yoichi Nakanishi and
Masayoshi Maeshima*
Laboratory of Biochemistry, Graduate School of Bioagricultural
Sciences, Nagoya University, Nagoya, 464-01 Japan (Y.N.,
M.M.); and Department of Cell Biology, National Institute for Basic
Biology, Okazaki, 444 Japan (M.M.)
Vacuolar proton-translocating
inorganic pyrophosphatase and H+-ATPase acidify the
vacuoles and power the vacuolar secondary active transport systems in
plants. Developmental changes in the transcription of the
pyrophosphatase in growing hypocotyls of mung bean (Vigna
radiata) were investigated. The cDNA clone for the mung bean
enzyme contains an uninterrupted open reading frame of 2298 bp, coding
for a polypeptide of 766 amino acids. Hypocotyls were divided into
elongating and mature regions. RNA analysis revealed that the
transcript level of the pyrophosphatase was high in the elongating
region of the 3-d-old hypocotyl but was extremely low in the mature
region of the 5-d-old hypocotyl. The level of transcript of the 68-kD
subunit of H+-ATPase also decreased after cell maturation.
In the elongating region, the proton-pumping activity of
pyrophosphatase on the basis of membrane protein was 3 times higher
than that of H+-ATPase. After cell maturation, the
pyrophosphatase activity decreased to 30% of that in the elongating
region. The decline in the pyrophosphatase activity was in parallel
with a decrease in the enzyme protein content. These findings indicate
that the level of the pyrophosphatase, a main vacuolar proton pump in
growing cells, is negatively regulated after cell maturation at the
transcriptional level.
1
Part of the research for this work was supported
by Grants-in-Aid for Scientific Research from the Ministry of
Education, Science, and Culture of Japan to M.M. (nos. 09257221 and
09660114).
*
Corresponding author; e-mail maeshima{at}agr.nagoya-u.ac.jp; fax
81-52-789-4094.
Plant Physiol. (1998) 116: 589-597
Copyright Clearance Center: 0032-0889/98/116/0589/09
© 1998 American Society of Plant Physiologists
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