Zhang L, Ohta A, Takagi M, Imai R
Department of Biotechnology, University of Tokyo, Yayoi, Japan.
J Biochem. 2000 Apr;127(4):611-6. doi: 10.1093/oxfordjournals.jbchem.a022648.
To study functions of late embryogenesis abundant (LEA) proteins, which accumulate in plant cells under water deficit conditions, in vivo functional analyses were carried out using a yeast (Saccharomyces cerevisiae) heterologous expression system. Two lea genes, tomato le4 (group 2) and barley HVA1 (group 3), were expressed under the GAL1 promoter, and the gene products were detected using specific antisera. The growth of the transformants was scored and compared with a control strain to analyze the effect of these proteins on yeast cells under stress conditions. The yeast cells expressing HVA1 showed shorter lag period when transferred to a medium containing 1.2 M NaCl as compared to a control strain, while the cells expressing le4 did not show improved growth. Attenuated growth inhibition in a medium containing 1.2 M KCl was observed in the yeast cells expressing le4 and HVA1. No obvious growth improvement was observed in a high sorbitol medium in the cells expressing either le4 or HVA1. Increased freezing tolerance was observed in both lea-expressing cells, while no effect on heat tolerance was observed. These results support the hypothesis that different LEA proteins play a distinctive role in the protection against cellular dehydration.
为了研究在水分亏缺条件下在植物细胞中积累的胚胎发育晚期丰富(LEA)蛋白的功能,利用酵母(酿酒酵母)异源表达系统进行了体内功能分析。两个lea基因,番茄le4(第2组)和大麦HVA1(第3组),在GAL1启动子下表达,并使用特异性抗血清检测基因产物。对转化体的生长进行评分,并与对照菌株进行比较,以分析这些蛋白质在胁迫条件下对酵母细胞的影响。与对照菌株相比,表达HVA1的酵母细胞转移到含有1.2 M NaCl的培养基中时滞后期较短,而表达le4的细胞生长没有改善。在表达le4和HVA1的酵母细胞中观察到在含有1.2 M KCl的培养基中生长抑制减弱。在表达le4或HVA1的细胞的高山梨醇培养基中未观察到明显的生长改善。在两个表达lea的细胞中均观察到冷冻耐受性增加,而未观察到对耐热性的影响。这些结果支持了不同的LEA蛋白在保护细胞免受脱水方面发挥独特作用的假设。