Abstract
Eukaryotic cells respond to the accumulation of unfolded proteins in the endoplasmic reticulum (ER). In this case, so-called unfolded protein response (UPR) genes are induced. We determined the transcriptional expression of Arabidopsis thaliana UPR genes by fluid microarray analysis of tunicamycin-treated plantlets. Two hundred and fifteen up-regulated genes and 17 down-regulated ones were identified. These genes were reanalyzed with functional DNA microarrays, using DNA fragments cloned through fluid microarray analysis. Finally, 36 up-regulated and two down-regulated genes were recognized as UPR genes. Among them, the up-regulation of genes related to protein degradation (HRD1, SEL-1L/HRD3 and DER1), regulation of translation (P58(IPK)), and apoptosis (BAX inhibitor-1) was reconfirmed by real-time reverse transcriptase-PCR. The induction of SEL-1L protein in an Arabidopsis membrane fraction on tunicamycin-treatment was demonstrated. Phosphorylation of initiation factor-2alpha, which was inhibited by P58(IPK), was decreased in tunicamycin-treated plantlets. However, regulatory changes in translation caused by ER stress were not detected in Arabidopsis. Plant cells appeared to have a strategy for overcoming ER stress through enhancement of protein folding activity, degradation of unfolded proteins, and regulation of apoptosis, but not regulation of translation.
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