Key Laboratory of Ministry of Education for Cell Proliferation and Differentiation, College of Life Sciences, Peking University, Beijing, China.
National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China.
J Exp Bot. 2018 Feb 23;69(5):1051-1064. doi: 10.1093/jxb/erx458.
The exocyst, an evolutionarily conserved octameric protein complex involved in exocytosis, has been reported to be involved in diverse aspects of morphogenesis in Arabidopsis. However, the molecular functions of such exocytotic molecules in rice are poorly understood. Here, we examined the molecular function of OsSEC3A, an important subunit of the exocyst complex in rice. The OsSEC3A gene is expressed in various organs, and OsSEC3A has the potential ability to participate in the exocyst complex by interacting with several other exocyst subunits. Disruption of OsSEC3A by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) caused dwarf stature and a lesion-mimic phenotype. The Ossec3a mutant exhibited enhanced defense responses, as shown by up-regulated transcript levels of pathogenesis- and salicylic acid synthesis-related genes, increased levels of salicylic acid, and enhanced resistance to the fungal pathogen Magnaporthe oryzae. Subcellular localization analysis demonstrated that OsSEC3A has a punctate distribution with the plasma membrane. In addition, OsSEC3A interacted with rice SNAP25-type t-SNARE protein OsSNAP32, which is involved in rice blast resistance, via the C-terminus and bound to phosphatidylinositol lipids, particularly phosphatidylinositol-3-phosphate, through its N-terminus. These findings uncover the novel function of rice exocyst subunit SEC3 in defense responses.
外泌体是一种进化上保守的八聚体蛋白复合物,参与细胞的胞吐作用,据报道它参与了拟南芥形态发生的多个方面。然而,水稻中这种胞吐分子的分子功能还知之甚少。在这里,我们研究了水稻外泌体复合物的重要亚基 OsSEC3A 的分子功能。OsSEC3A 基因在各种器官中表达,并且通过与其他几个外泌体亚基相互作用,OsSEC3A 具有参与外泌体复合物的潜力。CRISPR/Cas9(成簇规律间隔短回文重复/CRISPR 相关蛋白 9)敲除 OsSEC3A 导致矮化表型和损伤模拟表型。Ossec3a 突变体表现出增强的防御反应,表现为与发病机制和水杨酸合成相关基因的转录水平上调、水杨酸水平升高以及对真菌病原体稻瘟病菌的抗性增强。亚细胞定位分析表明,OsSEC3A 与质膜呈点状分布。此外,OsSEC3A 通过 C 末端与参与水稻稻瘟病抗性的水稻 SNAP25 型 t-SNARE 蛋白 OsSNAP32 相互作用,并通过其 N 末端与质膜结合,特别是与磷脂酰肌醇-3-磷酸结合。这些发现揭示了水稻外泌体亚基 SEC3 在防御反应中的新功能。