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.

摘要合作研究：解析RSH扩展蛋白在植物细胞壁组装中的作用植物细胞壁是一种自组装结构，其特性来源于正确的组装。细胞壁组装的故障可导致植物的早期死亡，正如拟南芥胚致死突变体所发生的那样，这是本提案的重点。rsh突变体不能在分裂细胞中组装功能细胞壁，因为它缺乏一种特定的富含羟基脯氨酸的糖蛋白，特别是一种名为rsh （ROOT-SHOOT-HYPOCOTYL-DEFECTIVE）的交联扩展蛋白。这种突变将使在分子水平上解剖壁自组装的精确规则成为可能。突变植株可以通过移植一系列新的RSH基因来治愈它们的缺陷，这些基因可能会取代缺失基因的功能。这些与一系列RSH类似物的互补实验将能够定义RSH对细胞壁组装的贡献。该策略包括操纵野生型基因和将通过合成基因产生的类似物。RSH是一种简单、高度重复的蛋白质，非常适合用于合成基因方法。某些氨基酸，包括赖氨酸、酪氨酸、丝氨酸和组氨酸，可能是组装RSH并帮助其交联形成植物细胞壁支架的关键，因此将产生基因，使RSH蛋白在这些氨基酸的含量上发生改变，这些基因治疗突变的能力将被评估。其他设计的RSH分子将缺乏蛋白质的一端（n端）或另一端（c端）的一部分，因为初步结果表明，RSH的一端（c端）是壁组装所必需的。最后，将对天然RSH、RSH类似物和拟南芥细胞壁进行生化表征。更广泛的影响：定义植物细胞壁中RSH的分子相互作用的长期目标，以及RSH中不同部分和氨基酸所起的确切作用，与了解细胞壁如何自组装有关，包括了解植物的防御反应和植物形态是如何形成的。更广泛的生物技术价值包括对超分子化学规则的新见解；设计能在分子水平上指导物质自组装的多用途分子是纳米技术的一个关键目标。这项工作还结合了细胞壁生物学和生物化学，以一种教导初级研究人员识别重要的生物学问题，并通过从蛋白质设计和表征到分子遗传学和细胞生物学的各种方法的合作，着手解决这个问题。两位研究人员都参加了与增加科学多样性有关的外展活动，包括向阿巴拉契亚地区外展。