Functional Domains and Spatial-Temporal Interactions of an Arabidopsis Circadian Clock-Associated F-Box Protein

拟南芥昼夜节律钟相关 F-Box 蛋白的功能域和时空相互作用

• 批准号: 0544137
• 负责人: David Somers
• 金额: --
• 依托单位: Ohio State University Research Foundation -DO NOT USE
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0544137&HistoricalAwards=false
• 关键词: Functional; Domains; Spatial; Temporal; Interactions

Ubiquitin-based proteolysis is a modification and degradation system that is essential to proper functioning of all eukaryotes. Of the cullin ubiquitin ligases, the SCF class is the most diverse, with nearly 700 F-box proteins identified. ZEITLUPE (ZTL) is a key F-box protein involved in regulating the plant circadian clock. The recent identification of a proteolytic target of ZTL (TOC1), will now allow focus on the functional dissection of ZTL, its intracellular site of action, and identify additional biochemical interactors. Using deletion constructs of ZTL, and newly identified point mutations, the domains and residues necessary for in planta TOC1/ZTL interactions will be identified. The spatial and temporal ZTL/TOC interaction will also be identified, by experimentally controlling and monitoring their compartmentation. Furthermore direct targets of ZTL-mediated degradation will be determined. The specific ASK family member(s) that participate in the native SCFZTL complex and other additional interacting partners will also be studied. This work will provide an opportunity for the graduate student and postdoctoral researchers involved to learn contemporary molecular biology and plant molecular genetic techniques in an interactive environment. Additionally, the more experienced lab workers will mentor undergraduates, so they will learn technical and molecular skills that they could use later in their career. Additional impacts include the development and instruction of a Circadian Biology course designed to survey and synthesize current topics in all circadian systems. The research experiences of the personnel in this project will broaden their scientific expertise, and contribute to their development as future PI's, post-docs and teachers.

基于泛素的蛋白水解是一种修饰和降解系统，对于所有真核生物的正常运作至关重要。在Cullin泛素连接酶中，SCF类是最多样化的，近700个F-box蛋白都鉴定出来。 Zeitlupe（ZTL）是用于调节植物昼夜节律时钟的关键F-box蛋白。最近对ZTL的蛋白水解靶标（TOC1）的鉴定将允许关注ZTL的功能解剖，即其细胞内作用部位，并确定其他生化相互作用者。使用ZTL的缺失构建体以及新鉴定的点突变，将确定Planta TOC1/ZTL相互作用所需的域和残基。还将通过实验控制和监视其隔室来确定空间和时间ZTL/TOC相互作用。此外，将确定ZTL介导的降解的直接靶标。还将研究参与本地SCFZTL综合体和其他其他互动伙伴的特定询问家庭成员。 这项工作将为参与研究生和博士后研究人员提供一个机会，以在交互式环境中学习当代分子生物学和植物分子遗传技术。此外，经验丰富的实验室工人将指导本科生，因此他们将学习技术和分子技能，在职业生涯中可以使用这些技能。其他影响包括昼夜节律生物学课程的开发和指导，旨在调查和综合所有昼夜节律系统中的当前主题。该项目中人员的研究经验将扩大其科学专业知识，并为他们作为未来的PI，dost-Docs和教师的发展做出贡献。