Browne John A, Dolan Katharine M, Tyson Trevor, Goyal Kshamata, Tunnacliffe Alan, Burnell Ann M
Institute of Bioengineering and Agroecology, Department of Biology, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland.
Eukaryot Cell. 2004 Aug;3(4):966-75. doi: 10.1128/EC.3.4.966-975.2004.
Some organisms can survive exposure to extreme desiccation by entering a state of suspended animation known as anhydrobiosis. The free-living nematode Aphelenchus avenae can be induced to enter the anhydrobiotic state by exposure to a moderate reduction in relative humidity. During this preconditioning period, the nematode accumulates large amounts of the disaccharide trehalose, which is thought to be necessary, but not sufficient, for successful anhydrobiosis. To identify other adaptations that are required for anhydrobiosis, we developed a novel SL1-based mRNA differential display technique to clone genes that are upregulated by dehydration in A. avenae. Three such genes, Aav-lea-1, Aav-ahn-1, and Aav-glx-1, encode, respectively, a late embryogenesis abundant (LEA) group 3 protein, a novel protein that we named anhydrin, and the antioxidant enzyme glutaredoxin. Strikingly, the predicted LEA and anhydrin proteins are highly hydrophilic and lack significant secondary structure in the hydrated state. The dehydration-induced upregulation of Aav-lea-1 and Aav-ahn-1 was confirmed by Northern hybridization and quantitative PCR experiments. Both genes were also upregulated by an osmotic upshift, but not by cold, heat, or oxidative stress. Experiments to investigate the relationship between mRNA levels and protein expression for these genes are in progress. LEA proteins occur commonly in plants, accumulating during seed maturation and desiccation stress; the presence of a gene encoding an LEA protein in an anhydrobiotic nematode suggests that some mechanisms of coping with water loss are conserved between plants and animals.
一些生物体可以通过进入一种称为脱水休眠的假死状态来在极端干燥环境下存活。自由生活的线虫燕麦真滑刃线虫可以通过暴露于相对湿度适度降低的环境中被诱导进入脱水休眠状态。在这个预处理阶段,线虫会积累大量的二糖海藻糖,人们认为海藻糖对于成功进入脱水休眠状态是必要的,但不是充分的。为了确定脱水休眠所需的其他适应性变化,我们开发了一种基于SL1的新型mRNA差异显示技术,以克隆在燕麦真滑刃线虫中因脱水而上调的基因。三个这样的基因,Aav-lea-1、Aav-ahn-1和Aav-glx-1,分别编码一种晚期胚胎发生丰富(LEA)3组蛋白、一种我们命名为脱水蛋白的新型蛋白质以及抗氧化酶谷氧还蛋白。引人注目的是,预测的LEA蛋白和脱水蛋白在水合状态下具有高度亲水性且缺乏明显的二级结构。通过Northern杂交和定量PCR实验证实了脱水诱导的Aav-lea-1和Aav-ahn-1上调。这两个基因也因渗透压升高而上调,但不因寒冷、高温或氧化应激而上调。研究这些基因的mRNA水平与蛋白质表达之间关系的实验正在进行中。LEA蛋白常见于植物中,在种子成熟和脱水胁迫期间积累;在脱水休眠线虫中存在编码LEA蛋白的基因表明,植物和动物之间应对水分流失的一些机制是保守的。