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THE PLANT CELL, Vol 9, Issue 8 1305-1316, Copyright © 1997 by American Society of Plant Biologists
Regulation of Lysine Catabolism through Lysine[mdash]Ketoglutarate Reductase and Saccharopine Dehydrogenase in Arabidopsis
G. Tang, D. Miron, J. X. Zhu-Shimoni and G. Galili
Department of Plant Genetics, Weizmann Institute of Science, Rehovot 76100, Israel
In plant and mammalian cells, excess lysine is catabolized by a pathway
that is initiated by two enzymes, namely, lysine-ketoglutarate reductase
and saccharopine dehydrogenase. In this study, we report the cloning of an
Arabidopsis cDNA encoding a bifunctional polypeptide that contains both of
these enzyme activities linked to each other. RNA gel blot analysis
identified two mRNA bands[mdash]a large mRNA containing both
lysine-ketoglutarate reductase and saccharopine dehydrogenase sequences and
a smaller mRNA containing only the saccharopine dehydrogenase sequence.
However, DNA gel blot hybridization using either the lysine-ketoglutarate
reductase or the saccharopine dehydrogenase cDNA sequence as a probe
suggested that the two mRNA populations apparently are encoded by the same
gene. To test whether these two mRNAs are functional, protein extracts from
Arabidopsis cells were fractionated by anion exchange chromatography. This
fractionation revealed two separate peaks[mdash]one containing both
coeluted lysine-ketoglutarate reductase and saccharopine dehydrogenase
activities and the second containing only saccharopine dehydrogenase
activity. RNA gel blot analysis and in situ hybridization showed that the
gene encoding lysine-ketoglutarate reductase and saccharopine dehydrogenase
is significantly upregulated in floral organs and in embryonic tissues of
developing seeds. Our results suggest that lysine catabolism is subject to
complex developmental and physiological regulation, which may operate at
gene expression as well as post-translational levels.
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