Molecular approaches towards control of sulfur flux in plants through selective deregulation of cysteine synthase complexes

通过选择性解除半胱氨酸合酶复合物控制植物硫通量的分子方法

• 批准号: 87777596
• 负责人: Professor Dr. Rüdiger Hell
• 金额: --
• 依托单位: Institute of Biochemistry and Biophysics
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/87777596?language=en
• 关键词: Molecular; approaches; towards; control; sulfur

Sulfur assimilation and cysteine biosynthesis in plants are part of the regulatory network of primary assimilation pathways that govern plant fitness and crop production. Protein complexes of cysteine synthase exist in three cellular compartments and are believed to act as regulators of primary sulfur metabolism via reversible protein-protein interaction. The aim of the project is a targeted deregulation of the formation of cysteine synthase complexes in specific compartments to study the consequences of such manipulations in planta. This will be achieved in three stages: An 'in silico' stage will include detailed analyses of the existing structural data of the two subunits of the cysteine synthase complex and the design of polypeptide candidates interfering with the interaction of subunits. During an 'in vitro' stage, the functionality of the designed peptides will be verified by biophysical and biochemical methods using recombinant purified forms of the subunits. The 'in vivo' stage will include transformation of tobacco with the pre-selected constructs and identification of genotypes with deregulated cysteine synthesis in mitochondria and chloroplasts. This will enable us to track down the structural basis of regulation within the cysteine synthase complex and its role in regulation of sulfur flux.

植物中硫的同化和半胱氨酸的生物合成是控制植物适合性和作物产量的初级同化途径调控网络的一部分。半胱氨酸合成酶的蛋白质复合体存在于三个细胞室，被认为是通过可逆的蛋白质-蛋白质相互作用来调节初级硫代谢的。该项目的目的是有针对性地放松对特定隔室中半胱氨酸合成酶复合体的形成的管制，以研究这种操纵在植物中的后果。这将分三个阶段实现：“在计算机”阶段将包括对半胱氨酸合成酶复合体两个亚基的现有结构数据的详细分析，以及干扰亚基相互作用的多肽候选的设计。在‘体外’阶段，设计的多肽的功能将通过生物物理和生化方法进行验证，方法是使用重组的亚基纯化形式。“体内”阶段将包括用预先选择的构建体转化烟草，以及鉴定线粒体和叶绿体中半胱氨酸合成失控的基因型。这将使我们能够追踪半胱氨酸合成酶复合体内调控的结构基础及其在硫通量调控中的作用。