A temperature-sensitive mechanism that regulates post-translational stability of a plastidial ω-3 fatty acid desaturase (FAD8) in Arabidopsis leaf tissues

Osamu Matsuda, Hikaru Sakamoto, Tadafumi Hashimoto, Koh Iba

Research output: Contribution to journalArticle

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Abstract

Trienoic fatty acids (TAs) are the major constituents in plant membrane lipids. In Arabidopsis, two plastidial isozymes of ω-3 fatty acid desaturase, FAD7 and FAD8, are the major contributors for TA production in leaf tissues. Despite a high degree of structural relatedness, activities of these two isozymes are regulated differentially in response to temperature. Elevated temperatures lead to decreases in leaf TA level due to temperature sensitivity of FAD8 activity. A series of FAD7-FAD8 chimeric genes, each encoding a functional plastidial ω-3 desaturase, were introduced into the Arabidopsis fad7fad8 double mutant. Constructs with or without a c-Myc epitope tag were tested. Functionality of each chimeric gene in response to temperature was assayed by Northern and Western analyses and by examining the fatty acid composition. All transformants harboring a chimeric gene containing the FAD8-derived C-terminal coding region (44 amino acids) showed a marked decrease in TA level when exposed to high temperature, similarly as transgenic lines complemented with the native form of FAD8. The reduction of TA level was accompanied by a decrease in the amount of ω-3 desaturase protein but not necessarily by a decrease in its transcript level. Analysis of the decay of c-Myc-tagged products after inhibiting protein synthesis revealed that the FAD8-derived C-terminal region acts in an autoregulatory fashion to destabilize the protein at high temperature. This suggests that the regulation of post-translational stability of FAD8 provides an important regulatory mechanism for modifying its activity in response to temperature, mediating a decrease in TA level at elevated temperatures.

Original languageEnglish
Pages (from-to)3597-3604
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number5
DOIs
Publication statusPublished - Feb 4 2005

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Fatty Acid Desaturases
Arabidopsis
Tissue
Fatty Acids
Temperature
Isoenzymes
Genes
Proteins
Gene encoding
Membrane Lipids
Epitopes
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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A temperature-sensitive mechanism that regulates post-translational stability of a plastidial ω-3 fatty acid desaturase (FAD8) in Arabidopsis leaf tissues. / Matsuda, Osamu; Sakamoto, Hikaru; Hashimoto, Tadafumi; Iba, Koh.

In: Journal of Biological Chemistry, Vol. 280, No. 5, 04.02.2005, p. 3597-3604.

Research output: Contribution to journalArticle

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