Kamauchi Shinya, Nakatani Hiromi, Nakano Chiharu, Urade Reiko
Graduate School of Agriculture, Kyoto University, Uji, Japan.
FEBS J. 2005 Jul;272(13):3461-76. doi: 10.1111/j.1742-4658.2005.04770.x.
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.
真核细胞会对内质网(ER)中未折叠蛋白的积累做出反应。在这种情况下,所谓的未折叠蛋白反应(UPR)基因会被诱导。我们通过对衣霉素处理的幼苗进行液相微阵列分析,确定了拟南芥UPR基因的转录表达。鉴定出215个上调基因和17个下调基因。使用通过液相微阵列分析克隆的DNA片段,用功能DNA微阵列对这些基因进行了重新分析。最后,36个上调基因和2个下调基因被确认为UPR基因。其中,通过实时逆转录聚合酶链反应再次证实了与蛋白质降解(HRD1、SEL-1L/HRD3和DER1)、翻译调控(P58(IPK))和细胞凋亡(BAX抑制剂-1)相关的基因上调。证明了衣霉素处理后拟南芥膜组分中SEL-1L蛋白的诱导。在衣霉素处理的幼苗中,被P58(IPK)抑制的起始因子-2α的磷酸化水平降低。然而,在拟南芥中未检测到内质网应激引起的翻译调控变化。植物细胞似乎有一种通过增强蛋白质折叠活性、降解未折叠蛋白和调控细胞凋亡来克服内质网应激的策略,而不是通过调控翻译。