State Key Laboratory for Crop Stress Resistance and High-Efficiency Production/Shaanxi Key Laboratory of Apple, College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China.
State Key Laboratory for Crop Stress Resistance and High-Efficiency Production/Shaanxi Key Laboratory of Apple, College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Gene. 2025 Jan 30;935:149076. doi: 10.1016/j.gene.2024.149076. Epub 2024 Nov 4.
E3 ligases are essential for ubiquitination and play a role in regulating various aspects of eukaryotic life. Ariadne (ARI) proteins, a subfamily of RBR (RING-between-RING) proteins, have been recognized as a new class of RING-finger E3 ligases. Recent research has shed light on their potential involvement in plants' responses to abiotic stress. However, comprehensive studies on ARI genes in apple (Malus domestica) are still lacking. This study identified ten MdARI genes in the apple genome, and examined intraspecific and interspecific collinearity to explore the evolutionary relationships of ARI family members. Phylogenetic analyses classified MdARIs into two subfamilies (A and B), and by integrating gene structure, conserved motifs, and sequence comparison results, subfamily B was further divided into two subgroups (I and II). Tissue expression analyses revealed varied expression patterns of MdARI genes in different tissues, and subcellular localization showed that MdARI1-1, MdARI1-2, and MdARI9-1 were located in the nucleus, while the other seven MdARIs were distributed throughout the cell. Analyses of promoter cis-elements and expression patterns under cold, salt, and drought treatments indicated the involvement of MdARIs in abiotic stress responses. Several proteins crucial to the plant stress response were predicted to be potential MdARIs-interacting proteins based on the protein interaction network. Additionally, the interaction between UBC11 (E2) and MdARI7-2 was confirmed by a yeast two-hybrid (Y2H) experiment, suggesting that MdARI7-2 may function as an E3. These findings will greatly benefit future research on the role and mechanisms of ARI proteins in apple stress response.
E3 连接酶对于泛素化至关重要，在调节真核生物生活的各个方面发挥作用。Ariadne（ARI）蛋白是 RBR（环指之间的环）蛋白的一个亚家族，已被认为是一类新的 RING 指 E3 连接酶。最近的研究揭示了它们在植物应对非生物胁迫中的潜在作用。然而，关于苹果（Malus domestica）中 ARI 基因的综合研究仍然缺乏。本研究在苹果基因组中鉴定了 10 个 MdARI 基因，并研究了种内和种间的共线性，以探讨 ARI 家族成员的进化关系。系统发育分析将 MdARIs 分为两个亚家族（A 和 B），通过整合基因结构、保守基序和序列比较结果，亚家族 B 进一步分为两个亚组（I 和 II）。组织表达分析显示 MdARI 基因在不同组织中的表达模式存在差异，亚细胞定位表明 MdARI1-1、MdARI1-2 和 MdARI9-1 位于细胞核中，而其他 7 个 MdARIs 则分布在整个细胞中。对启动子顺式元件和冷、盐、干旱处理下的表达模式分析表明，MdARIs 参与了非生物胁迫反应。根据蛋白质相互作用网络，预测了几种对植物应激反应至关重要的蛋白质可能是潜在的 MdARIs 相互作用蛋白。此外，通过酵母双杂交（Y2H）实验证实了 UBC11（E2）和 MdARI7-2 之间的相互作用，表明 MdARI7-2 可能作为 E3 发挥作用。这些发现将极大地促进未来对 ARI 蛋白在苹果应激反应中的作用和机制的研究。
