New Twists in CDPK Regulation: dual-specificity kinases and a role for calmodulin

CDPK 调节的新转折：双特异性激酶和钙调蛋白的作用

• 批准号: 1354094
• 负责人: Raymond Zielinski
• 金额: $ 53.53万
• 依托单位: University of Illinois at Urbana-Champaign
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1354094&HistoricalAwards=false
• 关键词: New; Twists; CDPK; Regulation; dual

Plant calcium-dependent protein kinases (CPKs or CDPKs) are a large family of plant protein kinases that act as calcium sensors to regulate a range of processes involved in regulation of growth, stomatal aperture, hormone signaling, and immune/stress signaling. Hence, a solid understanding of the mechanisms that regulate these protein kinases is of fundamental importance. The aim of this project is to further elucidate two recently discovered novel properties of CDPKs, tyrosine autophosphorylation and calmodulin (CaM)-binding, to better understand how these factors regulate CDPK activity and plant function. The overall goals of the project are to understand how posttranslational modifications of specific CDPKs modulates their function, and to define the role these modifications play in regulating plant growth, development and responses to environmental stimuli. The approach will involve generating and measuring the activities of phosphosite CDPK mutants, determining their effect on CaM-binding, and using the directed mutants to complement CDPK null mutants to determine their effect on plant phenotype. The project will center around two broad objectives to characterize the occurrence and regulatory role of: 1) autophosphorylation of CDPKs on Ser, Thr and Tyr residues, and 2) CaM binding to CDPKs. Studies will involve recombinant proteins (and site-directed mutants) in vitro that will guide transgenic approaches to examine function in planta. The results of the proposed studies may fundamentally alter two paradigms of CDPK regulation. In addition to providing traditional student training, the project will also include summer programs that provide opportunities for recruiting underrepresented groups and involve students in lab activities. In addition, the new Professional Science Master's Program in Plant Biology/Biotechnology will engage students who are training for positions outside of academic research with a non-thesis research project. Finally, all biological materials derived from the project will be made available to the research community.

植物钙依赖性蛋白激酶（CPK 或 CDPK）是植物蛋白激酶的一个大家族，充当钙传感器来调节涉及生长、气孔孔径、激素信号传导和免疫/应激信号传导的一系列过程。 因此，深入了解调节这些蛋白激酶的机制至关重要。 该项目的目的是进一步阐明最近发现的 CDPK 的两个新特性：酪氨酸自磷酸化和钙调蛋白 (CaM) 结合，以更好地了解这些因素如何调节 CDPK 活性和植物功能。该项目的总体目标是了解特定 CDPK 的翻译后修饰如何调节其功能，并确定这些修饰在调节植物生长、发育和对环境刺激的反应中所发挥的作用。 该方法将涉及生成和测量磷酸位 CDPK 突变体的活性，确定它们对 CaM 结合的影响，并使用定向突变体补充 CDPK 无效突变体以确定它们对植物表型的影响。 该项目将围绕两个广泛的目标来表征以下方面的发生和调节作用：1) CDPK 在 Ser、Thr 和 Tyr 残基上的自磷酸化，以及 2) CaM 与 CDPK 的结合。 研究将涉及体外重组蛋白（和定点突变体），这将指导转基因方法检查植物中的功能。 拟议研究的结果可能从根本上改变 CDPK 调节的两种范式。除了提供传统的学生培训外，该项目还将包括暑期项目，为招募代表性不足的群体提供机会，并让学生参与实验室活动。 此外，新的植物生物学/生物技术专业科学硕士课程将让正在接受学术研究以外职位培训的学生参与非论文研究项目。 最后，来自该项目的所有生物材料将提供给研究界。