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THE PLANT CELL, Vol 8, Issue 7 1159-1169, Copyright © 1996 by American Society of Plant Biologists
A Single Gene May Encode Differentially Localized Ca2+-ATPases in Tomato
N. Ferrol and A. B. Bennett
Mann Laboratory, Department of Vegetable Crops, University of California at Davis, Davis, California 95616
Previously, a partial-length cDNA and a complete genomic clone encoding a
putative sarcoplasmic reticulum-type Ca2+-ATPase (LCA, Lycopersicon
Ca2+-ATPase) were isolated from tomato. To determine the subcellular
localization of this Ca2+-ATPase, specific polyclonal antibodies raised
against a fusion protein encoding a portion of the LCA polypeptide were
generated. Based on hybridization of the LCA cDNA and of the nucleotide
sequence encoding the fusion protein to genomic DNA, it appears that LCA
and the fusion protein domain are encoded by a single gene in tomato.
Antibodies raised against the LCA domain fusion protein reacted
specifically with two polypeptides of 116 and 120 kD that are localized in
the vacuolar and plasma membranes, respectively. The distribution of
vanadate-sensitive ATP-dependent Ca2+ transport activities in sucrose
gradients coincided with the distribution of the immunodetected proteins.
The ATP-dependent Ca2+ transport activities associated with tonoplast and
plasma membrane fractions shared similar properties, because both fractions
were inhibited by vanadate but insensitive to carbonyl cyanide
m-chlorophenylhydrazone, nitrate, and calmodulin. Moreover, antibodies
raised against the LCA domain fusion protein inhibited ATP-dependent Ca2+
uptake activity associated with both the tonoplast and plasma membrane
fractions. These data suggest that a single gene (LCA) may encode two
P-type Ca2+-ATPase isoforms that are differentially localized in the
tonoplast and plasma membrane of tomato roots.
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