Proteome Analysis of Detergent-Resistant Membranes (DRMs) Associated with OsRac1-Mediated Innate Immunity in Rice
Proteome Analysis of Detergent-Resistant Membranes (DRMs) Associated with OsRac1-Mediated Innate Immunity in Rice
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DOI：
10.1093/pcp/pcp077
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发表时间：
2009-07-01
影响因子：
4.9
通讯作者：
Shimamoto, Ko
中科院分区：
文献类型：
--
作者：
Fujiwara, Masayuki;Hamada, Satoshi;Shimamoto, Ko
OsRac1 是 Rac/Rop GTPase 家族的成员，作为分子开关在水稻先天免疫中发挥着重要作用，OsRac1 的活性形式在质膜 (PM) 中发挥作用。为了研究 OsRac1 在 PM 中的精确定位及其与其他信号成分的可能关联，我们对从用 myc 标记的组成型活性 (CA) OsRac1 转化的水稻悬浮培养细胞中分离出的 DRM（耐洗涤剂膜）进行了蛋白质组学分析。 DRM 是在寒冷条件下 Triton X-100 处理后不溶解的 PM 区域，被认为参与动物、酵母和植物细胞中的各种信号传导过程。我们在 DRM 中鉴定了 192 种蛋白质，其中包括受体样激酶 (RLK)，如 Xa21、核苷酸结合富含亮氨酸重复序列 (NB-LRR) 型抗病蛋白、糖基磷脂酰肌醇 (GPI) 锚定蛋白、突触融合蛋白、NADPH 氧化酶、WD-40 重复家族蛋白和各种 GTP 结合蛋白。其中许多蛋白质之前已在从其他植物物种、动物和酵母细胞中分离的 DRM 中被鉴定出，验证了我们研究中使用的方法。为了研究 DRM 和 OsRac1 介导的先天免疫之间可能的关联，我们使用了用 myc 标记的野生型 (WT) OsRac1 转化的水稻悬浮培养细胞，发现 OsRac1 和 OsRac1 的效应子 RACK1A 在几丁质引发剂处理后转移到 DRM。这些结果表明 OsRac1 介导的先天免疫与 PM 中的 DRM 相关。
OsRac1, a member of the Rac/Rop GTPase family, plays important roles as a molecular switch in rice innate immunity, and the active form of OsRac1 functions in the plasma membrane (PM). To study the precise localization of OsRac1 in the PM and its possible association with other signaling components, we performed proteomic analysis of DRMs (detergent-resistant membranes) isolated from rice suspension-cultured cells transformed with myc-tagged constitutively active (CA) OsRac1. DRMs are regions of the PM that are insoluble after Triton X-100 treatment under cold conditions and are thought to be involved in various signaling processes in animal, yeast and plant cells. We identified 192 proteins in DRMs that included receptor-like kinases (RLKs) such as Xa21, nucleotide-binding leucine-rich repeat (NB-LRR)-type disease resistance proteins, a glycosylphosphatidylinositol (GPI)-anchored protein, syntaxin, NADPH oxidase, a WD-40 repeat family protein and various GTP-binding proteins. Many of these proteins have been previously identified in the DRMs isolated from other plant species, and animal and yeast cells, validating the methods used in our study. To examine the possible association of DRMs and OsRac1-mediated innate immunity, we used rice suspension-cultured cells transformed with myc-tagged wild-type (WT) OsRac1 and found that OsRac1 and RACK1A, an effector of OsRac1, shifted to the DRMs after chitin elicitor treatment. These results suggest that OsRac1-mediated innate immunity is associated with DRMs in the PM.