Danyluk J, Perron A, Houde M, Limin A, Fowler B, Benhamou N, Sarhan F
Departement des Sciences Biologiques, Universite du Quebec a Montreal, C.P. 8888 Succursale Centre-ville, Montreal, Quebec H3C 3P8, Canada.
Plant Cell. 1998 Apr;10(4):623-38. doi: 10.1105/tpc.10.4.623.
Expression of the acidic dehydrin gene wcor410 was found to be associated with the development of freezing tolerance in several Gramineae species. This gene is part of a family of three homologous members, wcor410, wcor410b, and wcor410c, that have been mapped to the long arms of the homologous group 6 chromosomes of hexaploid wheat. To gain insight into the function of this gene family, antibodies were raised against the WCOR410 protein and affinity purified to eliminate cross-reactivity with the WCS120 dehydrin-like protein of wheat. Protein gel blot analyses showed that the accumulation of WCOR410 proteins correlates well with the capacity of each cultivar to cold acclimate and develop freezing tolerance. Immunoelectron microscope analyses revealed that these proteins accumulate in the vicinity of the plasma membrane of cells in the sensitive vascular transition area where freeze-induced dehydration is likely to be more severe. Biochemical fractionation experiments indicated that WCOR410 is a peripheral protein and not an integral membrane protein. These results provide direct evidence that a subtype of the dehydrin family accumulates near the plasma membrane. The properties, abundance, and localization of these proteins suggest that they are involved in the cryoprotection of the plasma membrane against freezing or dehydration stress. We propose that WCOR410 plays a role in preventing the destabilization of the plasma membrane that occurs during dehydrative conditions.
酸性脱水素基因wcor410的表达与几种禾本科植物的抗冻性发育有关。该基因是由wcor410、wcor410b和wcor410c三个同源成员组成的基因家族的一部分,这些成员已被定位到六倍体小麦6号同源染色体组的长臂上。为了深入了解这个基因家族的功能,制备了针对WCOR410蛋白的抗体,并进行亲和纯化以消除与小麦WCS120脱水素样蛋白的交叉反应。蛋白质凝胶印迹分析表明,WCOR410蛋白的积累与每个品种的冷驯化和抗冻能力密切相关。免疫电子显微镜分析显示,这些蛋白质积累在敏感维管束过渡区细胞的质膜附近,在那里冷冻诱导的脱水可能更严重。生化分级实验表明,WCOR410是一种外周蛋白,而不是整合膜蛋白。这些结果提供了直接证据,表明脱水素家族的一个亚型在质膜附近积累。这些蛋白质的特性、丰度和定位表明它们参与了质膜对冷冻或脱水胁迫的低温保护。我们认为WCOR410在防止脱水条件下发生的质膜不稳定方面发挥作用。