Collaborative Research: Dissecting the Role of RSH Extensin in Assembly of the Plant Cell Wall

合作研究：剖析 RSH 延伸蛋白在植物细胞壁组装中的作用

• 批准号: 0622394
• 负责人: Marcia Kieliszewski
• 金额: --
• 依托单位: Ohio University
• 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: 2006-08-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0622394&HistoricalAwards=false
• 关键词: Collaborative; Research; Dissecting; Role; RSH

Kieliszewski/Cannon Proposal AbstractCOLLABORATIVE RESEARCH: Dissecting the role of RSH extensinin assembly of the plant cell wallThe plant cell wall is a self-assembling structure whose properties arise from correct assembly. Malfunctions in wall assembly can result in the early death of the plant, as occurs with the Arabidopsis rsh embryo lethal mutant, the focus of this proposal. The rsh mutant cannot assemble a functional cell wall in dividing cells because it lacks a specific hydroxyproline-rich glycoprotein, notably a crosslinking extensin designated RSH (ROOT-SHOOT-HYPOCOTYL-DEFECTIVE). This mutant will enable dissection of the precise rules for wall self-assembly at a molecular level. Mutant plants will be cured of their defect by transferring a range of new RSH genes that might replace the function of the gene that is missing. These complementation experiments with a series of RSH analogs will enable definition of the contributions RSH makes to cell wall assembly. The strategy involves manipulations of the wild-type gene and analogs that will be produced through synthetic genes. RSH is ideally suited for a synthetic gene approach as it is a simple, highly repetitive protein. Certain amino acids, including lysine, tyrosine, serine and histidine may be crucial to assemble RSH and help it crosslink it to form a scaffold in the plant cell wall, therefore genes will be made that give rise to RSH proteins altered in their content of these amino acids and the ability of these genes to cure the mutant will be evaluated. Other designer RSH molecules will lack part of one extreme end of the protein (N-terminus) or the other (C-terminus) as preliminary results indicate one end in particular, the C-terminus of RSH, is required for wall assembly. Finally naturally-occurring RSH, the RSH analogs and Arabidopsis cell walls will be characterized biochemically.Broader Impact: The long term goal of defining the molecular interactions of RSH in the plant cell wall, and the precise roles played by the different sections and amino acids in RSH are relevant to understanding how the wall self-assembles, including understanding plant defense responses and how plant form is created. A broader biotechnological value includes new insights into the rules of supramolecular chemistry; the design of versatile molecules that direct the self-assembly of matter at a molecular level is a key goal of nanotechnology. This work also integrates cell wall biology and biochemistry in a way that teaches junior researchers to identify a significant biological problem and set about solving it through collaboration that integrates methods ranging from protein design and characterization to molecular genetics and cell biology. Both investigators take part in outreach activities relevant to increasing diversity in the sciences, including outreach to Appalachia.

Kieliszewski/Cannon提案摘要Collaborative研究：解剖植物细胞壁的RSH延伸素组装的作用植物细胞壁是一种自组装的结构，其特性由正确的组装产生。 壁组件中的故障可能会导致植物的早期死亡，就像拟南芥RSH胚胎致命突变体发生的那样，这是该提案的重点。 RSH突变体无法在分裂细胞中组装功能性细胞壁，因为它缺乏特定的富含羟化糖蛋白，尤其是一种指定的RSH（根 - - 霍托赛 - 核基化）的交联延伸链蛋白。该突变体将在分子水平上解剖墙自组装的精确规则。突变植物将通过转移可能取代缺失基因功能的一系列新的RSH基因来治愈其缺陷。这些与一系列RSH类似物的互补实验可以定义RSH对细胞壁组装的贡献。该策略涉及通过合成基因产生的野生型基因和类似物的操纵。 RSH非常适合合成基因方法，因为它是一种简单的，高度重复的蛋白质。 某些氨基酸，包括赖氨酸，酪氨酸，丝氨酸和组氨酸，可能对组装RSH组装并帮助其交联以在植物细胞壁中形成支架，因此将制作基因，从而导致这些氨基酸含量改变的RSH蛋白质以及这些氨基酸的含量以及这些突变的能力可以评估这些氨基酸的能力。 其他设计师的RSH分子将缺少蛋白质（N末端）或另一个（C-末端）的一个极端的一部分，因为初步结果尤其表明一个末端，RSH的C末端是壁组件所必需的。最后，自然存在的RSH，RSH类似物和拟南芥细胞壁将被表征生化。Broader的影响：定义植物细胞壁中RSH分子相互作用的长期目标，以及不同部分和氨基酸在RSH中所起的精确作用与RSH中的不同型号和氨基酸相关，与理解壁上的植物构成了如何理解工厂的辩护，以及如何理解IS wayses，以及如何理解IS的辩护。更广泛的生物技术价值包括对超分子化学规则的新见解；将物质自组装在分子水平上的多功能分子的设计是纳米技术的关键目标。这项工作还以一种教会初级研究人员确定重要的生物学问题的方式整合细胞壁生物学和生物化学，并通过协作来解决它，从而整合从蛋白质设计和表征到分子遗传学和细胞生物学的方法。 两位调查人员都参加了与不断增加科学多样性有关的外展活动，包括向阿巴拉契亚宣讲。