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PLANT PHYSIOLOGY , Vol 113, Issue 4 1379-1384, Copyright © 1997 by American Society of Plant Biologists
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BIOCHEMISTRY AND ENZYMOLOGY |
Divergent Fructokinase Genes Are Differentially Expressed in Tomato
Y. Kanayama, N. Dai, D. Granot, M. Petreikov, A. Schaffer and A. B. Bennett
Mann Laboratory, Department of Vegetable Crops, University of California, Davis, California 95616 (Y.K., A.B.B.)
Two cDNA clones (Frk1 and Frk2) encoding fructokinase (EC 2.7.1.4) were
isolated from tomato (Lycopersicon esculentum). The Frk2 cDNA encoded a
deduced protein of 328 amino acids that was more than 90% identical with a
previously characterized potato (Solanum tuberosum) fructokinase. In
contrast, the Frk1 cDNA encoded a deduced protein of 347 amino acids that
shared only 55% amino acid identity with Frk2. Both deduced proteins
possessed an ATP-binding motif and putative substrate recognition site
sequences identified in bacterial fructokinases. The Frk1 cDNA was
expressed in a mutant yeast (Saccharomyces cerevisiae) line, which lacks
the ability to phosphorylate glucose and fructose and is unable to grow on
glucose or fructose. Mutant cells expressing Frk1 were complemented to grow
on fructose but not glucose, indicating that Frk1 phosphorylates fructose
but not glucose, and this activity was verified in extracts of transformed
yeast. The mRNA corresponding to Frk2 accumulated to high levels in young,
developing tomato fruit, whereas the Frk1 mRNA accumulated to higher levels
late in fruit development. The results indicate that fructokinase in tomato
is encoded by two divergent genes, which exhibit a differential pattern of
expression during fruit development.
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