The role of CDPKs in calcium sensing and ABA signaling

CDPK 在钙传感和 ABA 信号传导中的作用

• 批准号: 6790273
• 负责人: YINGZHEN YANG
• 金额: $ 4.3万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6790273
• 关键词: Arabidopsis; affinity chromatography; biological signal transduction; calcium flux; enzyme substrate; fluorescence microscopy; hormone regulation /control mechanism; immunoelectron microscopy; mass spectrometry; phytohormones; plant physiology; postdoctoral investigator; protein kinase; protein localization; yeast two hybrid system

DESCRIPTION (provided by applicant): Calcium-dependent protein kinases (CDPKs) constitute a specific protein kinase family containing both a kinase domain and a Ca2+ binding domain. There are 34 CDPKs encoded in Arabidopsis genome, but no phenotypes have been published for any loss-of-function cdpk mutants. The preliminary results of the sponsor's lab suggest that double mutants in two guard cell expressed CDPKs (CDPK-GCs) exhibit defects in ABA-induced stomatal closure. The phenotype of this cdpk-gc double mutant in ABA signaling provides a genetic tool to analyze the role of CDPKs in Ca2+ sensing and ABA signaling. [Ca2+]cyt oscillations are essential for long-term stomatal closure. The relative location of CDPK-GCs to [Ca2+]cyt oscillations in ABA signaling pathway will be determined by calcium imaging and stomatal movement analyses. The subcellular localization of CDPK-GCs will be determined by confocal fluorescent microscopy and immunogold electron microscopy. Substrates and interacting proteins of CDPK-GCs will be isolated by yeast two-hybrid assay and tandem affinity purification/mass spectrometry. The long-term goal is to use tagged CDPK substrates to test the hypothesis that CDPKs function as decoders of [Ca2+]cyt oscillations. Understanding the decoding mechanism of [Ca2+]cyt oscillation will contribute to the broad field of Ca2+ signaling including memory study in human.

描述（由申请人提供）： 钙依赖性蛋白激酶 (CDPK) 构成一个特定的蛋白激酶家族，包含激酶结构域和 Ca2+ 结合结构域。拟南芥基因组中编码了 34 个 CDPK，但尚未公布任何功能丧失的 cdpk 突变体的表型。申办者实验室的初步结果表明，两个保卫细胞表达的 CDPK (CDPK-GC) 中的双突变体在 ABA 诱导的气孔关闭中表现出缺陷。这种 cdpk-gc 双突变体在 ABA 信号传导中的表型为分析 CDPK 在 Ca2+ 传感和 ABA 信号传导中的作用提供了遗传工具。 [Ca2+]细胞色素振荡对于长期气孔关闭至关重要。 CDPK-GCs 与 ABA 信号通路中 [Ca2+]cyt 振荡的相对位置将通过钙成像和气孔运动分析来确定。 CDPK-GC 的亚细胞定位将通过共焦荧光显微镜和免疫金电子显微镜确定。 CDPK-GC 的底物和相互作用蛋白将通过酵母双杂交测定和串联亲和纯化/质谱法分离。长期目标是使用标记的 CDPK 底物来检验 CDPK 作为 [Ca2+]cyt 振荡解码器的假设。了解 [Ca2+]cyt 振荡的解码机制将有助于 Ca2+ 信号传导的广泛领域，包括人类记忆研究。