Department of Applied Plant Sciences Technology, Kangwon National University, Chuncheon 200-713, Republic of Korea.
DNA Res. 2013 Jun;20(3):299-314. doi: 10.1093/dnares/dst011. Epub 2013 Apr 9.
A large number of really interesting new gene (RING) E3 ligases contribute to the post-translational modification of target proteins during plant responses to environmental stresses. However, the physical interactome of RING E3 ligases in rice remains largely unknown. Here, we evaluated the expression patterns of 47 Oryza sativa RING finger protein (OsRFP) genes in response to abiotic stresses via semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and in silico analysis. Subsequently, molecular dissection of nine OsRFPs was performed by the examination of their E3 ubiquitin ligase activity, subcellular localization, and physical interaction with target proteins. Most of the OsRFPs examined possessed E3 ligase activity and showed diverse subcellular localization. Yeast two-hybrid analysis was then employed to construct a physical interaction map of seven OsRFPs with their 120 interacting proteins. The results indicated that these OsRFPs required dynamic translocation and partitioning for their cellular activation. Heterogeneous overexpression of each of the OsRFP genes in Arabidopsis suggested that they have functionally diverse responses to abiotic stresses, which may have been acquired during evolution. This comprehensive study provides insights into the biological functions of OsRFPs, which may be useful in understanding how rice plants adapt to unfavourable environmental conditions.
大量新的真核生物(RING)E3 连接酶在植物应对环境胁迫时参与靶蛋白的翻译后修饰。然而,水稻中 RING E3 连接酶的物理相互作用组在很大程度上仍是未知的。在这里,我们通过半定量逆转录聚合酶链反应(RT-PCR)和计算机分析评估了 47 个水稻 RING 指蛋白(OsRFP)基因对非生物胁迫的表达模式。随后,通过对 9 个 OsRFPs 的 E3 泛素连接酶活性、亚细胞定位和与靶蛋白的物理相互作用进行分子剖析。大多数受检的 OsRFPs 具有 E3 连接酶活性,并表现出不同的亚细胞定位。然后,利用酵母双杂交分析构建了 7 个 OsRFPs 与其 120 个相互作用蛋白的物理相互作用图谱。结果表明,这些 OsRFPs 为了细胞激活需要动态易位和分区。拟南芥中每个 OsRFP 基因的异源过表达表明,它们对非生物胁迫具有功能多样的响应,这可能是在进化过程中获得的。这项全面的研究提供了对 OsRFPs 生物学功能的深入了解,这可能有助于理解水稻植物如何适应不利的环境条件。