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植物-病原体相互作用中的 unfoldome 变化:草莓炭疽病菌感染。

Unfoldome variation upon plant-pathogen interactions: strawberry infection by Colletotrichum acutatum.

机构信息

Department of Agricultural Sciences, University of Bologna, Bologna, Italy.

Research and Innovation Centre, Foundation Edmund Mach (FEM), San Michele all' Adige, Trento, Italy.

出版信息

Plant Mol Biol. 2015 Sep;89(1-2):49-65. doi: 10.1007/s11103-015-0353-7. Epub 2015 Aug 6.

Abstract

Intrinsically disordered proteins (IDPs) are proteins that lack secondary and/or tertiary structure under physiological conditions. These proteins are very abundant in eukaryotic proteomes and play crucial roles in all molecular mechanisms underlying the response to environmental challenges. In plants, different IDPs involved in stress response have been identified and characterized. Nevertheless, a comprehensive evaluation of protein disorder in plant proteomes under abiotic or biotic stresses is not available so far. In the present work the transcriptome dataset of strawberry (Fragaria X ananassa) fruits interacting with the fungal pathogen Colletotrichum acutatum was actualized onto the woodland strawberry (Fragaria vesca) genome. The obtained cDNA sequences were translated into protein sequences, which were subsequently subjected to disorder analysis. The results, providing the first estimation of disorder abundance associated to plant infection, showed that the proteome activated in the strawberry red fruit during the active fungal propagation is remarkably depleted in disorder. On the other hand, in the resistant white fruit, no significant disorder reduction is observed in the proteins expressed in response to fungal infection. Four representative proteins, FvSMP, FvPRKRIP, FvPCD-4 and FvFAM32A-like, predicted as mainly disordered and never experimentally characterized before, were isolated, and the absence of structure was validated at the secondary and tertiary level using circular dichroism and differential scanning fluorimetry. Their quaternary structure was also established using light scattering. The results are discussed considering the role of protein disorder in plant defense.

摘要

无规卷曲蛋白质(IDPs)是指在生理条件下缺乏二级和/或三级结构的蛋白质。这些蛋白质在真核生物蛋白质组中非常丰富,在响应环境挑战的所有分子机制中发挥着关键作用。在植物中,已经鉴定和描述了不同参与应激反应的 IDP。然而,目前还没有针对非生物或生物胁迫下植物蛋白质组中蛋白质无序性的全面评估。在本工作中,将草莓( Fragaria X ananassa )果实与真菌病原体胶孢炭疽菌相互作用的转录组数据集实际应用于林地草莓( Fragaria vesca )基因组。获得的 cDNA 序列被翻译成蛋白质序列,随后对这些蛋白质序列进行无序分析。结果提供了与植物感染相关的无序丰度的首次估计,表明在草莓红色果实中真菌繁殖活跃期间激活的蛋白质组显著缺乏无序性。另一方面,在抗性白色果实中,真菌感染诱导表达的蛋白质中没有观察到无序性的显著减少。预测为主要无规卷曲且以前从未进行过实验表征的四个代表性蛋白质 FvSMP、FvPRKRIP、FvPCD-4 和 FvFAM32A-like 被分离出来,并使用圆二色性和差示扫描荧光法在二级和三级水平上验证了它们没有结构。还使用光散射法确定了它们的四级结构。考虑到蛋白质无序在植物防御中的作用,对结果进行了讨论。

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