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Biosynthesis of Lipoic Acid in Arabidopsis:
Cloning and Characterization of the cDNA for Lipoic Acid
Synthase1
Rie Yasuno and
Hajime Wada*
Department of Biology, Faculty of Science, Kyushu University,
Ropponmatsu, Fukuoka 810-8560, Japan
Lipoic acid is a coenzyme that is
essential for the activity of enzyme complexes such as those of
pyruvate dehydrogenase and glycine decarboxylase. We report here the
isolation and characterization of LIP1 cDNA for lipoic
acid synthase of Arabidopsis. The Arabidopsis LIP1 cDNA was isolated using an expressed sequence tag
homologous to the lipoic acid synthase of Escherichia
coli. This cDNA was shown to code for
Arabidopsis lipoic acid synthase by its ability to complement a
lipA mutant of E. coli defective in
lipoic acid synthase. DNA-sequence analysis of the LIP1
cDNA revealed an open reading frame predicting a protein of 374 amino
acids. Comparisons of the deduced amino acid sequence with those of
E. coli and yeast lipoic acid synthase homologs showed a
high degree of sequence similarity and the presence of a leader
sequence presumably required for import into the mitochondria.
Southern-hybridization analysis suggested that LIP1 is a
single-copy gene in Arabidopsis. Western analysis with an antibody
against lipoic acid synthase demonstrated that this enzyme is located
in the mitochondrial compartment in Arabidopsis cells as a 43-kD
polypeptide.
1
This work was supported by a Grant-in-Aid for
Scientific Research (no. 10640636) from the Ministry of Education,
Science, Sports, and Culture (Japan) and by a grant from the Asahi
Glass Foundation to H.W.
*
Corresponding author; e-mail wadarcb{at}mbox.nc.kyushu-u.ac.jp; fax
81-92-726-4761.
Plant Physiol. (1998) 118: 935-943
Copyright Clearance Center: 0032-0889/98/118//09
© 1998 American Society of Plant Physiologists
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