CRYSTAL STRUCTURE OF THE PLANT FULL-LENGTH SSDNA BINDING PROTEIN STWHY2 IN FREE

免费植物全长 SSDNA 结合蛋白 STWHY2 的晶体结构

• 批准号: 8363407
• 负责人: JURGEN SYGUSCH
• 金额: $ 0.69万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363407
• 关键词: Binding; Binding Proteins; C-terminal; Characteristics; Complex; Crystallization; Crystallography; DNA; Data; Funding; Gene Family; Genes; Genetic; Genetic Transcription; Grant; Knock-out; Knowledge; Lead; Length; Light; Location; Molecular Conformation; National Center for Research Resources; Phenotype; Plant Components; Plant Genes; Plants; Play; Potato; Principal Investigator; Protein Family; Proteins; Research; Research Infrastructure; Resources; Role; Source; Synchrotrons; United States National Institutes of Health; cost; defense response; design; member; novel; pathogen; promoter; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The plant defense response to pathogens relies primarily on the use of pre-existing physical barriers and inducible defenses, which require massive transcription of defense genes. The whirly protein family was discovered when studying the activation of such defense gene. StWhy1 was found as the activator of the potato plant gene PR-10a that is activated during defense response by binding to the promoter in its single stranded conformation. The protein was later crystallized and revealed novel tetramer whirligig-like¿ folding. As we tried to move forward in identifying the mechanism used by the members of this gene family, we have encountered difficulties taking the genetic approach. The phenotype shown by various Knockout of those genes are rare and subtle. Since we suspect the C-terminal part of those proteins to play big role in the Whirly function, we would like to obtain structural data of this region. Since the StWhy1 construct used then was not full length and it had a tag in place of C-terminal end, we need to use new construction. A paralogue of StWhy1, StWhy2, show a great potential for crystallization and hope to use this characteristic to obtain good crystals of the free form and the form in complex with ssDNA. We hope that those constructions will help reveal the location and potential role of the C-terminal domain when the protein is free or bound to DNA. This knowledge will guide our design of new experiments in the lab and lead us to better understanding of this protein family that plays an important role in plant defense against pathogens.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供 该子项目的主要支持。 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源的子项目可能列出的总成本。 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 植物对病原体的防御反应主要依赖于使用预先存在的物理屏障和诱导防御，这需要防御基因的大量转录，在研究此类防御基因的激活时发现了旋转蛋白家族。马铃薯植物基因 PR-10a 在防御反应期间通过与其单链构象的启动子结合而被激活。该蛋白质随后结晶并显示出新型四聚体旋转样。当我们试图进一步确定该基因家族成员所使用的机制时，我们在采用遗传方法时遇到了困难。这些基因的各种敲除所显示的表型是罕见且微妙的。这些蛋白质的末端部分在 Whirly 功能中发挥重要作用，我们希望获得该区域的结构数据，因为当时使用的 StWhy1 构建体不是全长，并且它有一个标签代替 C 末端，我们需要。使用新建筑。 StWhy1、StWhy2显示出巨大的结晶潜力，希望利用这一特性很好地获得游离形式和与ssDNA复合物形式的晶体，我们希望这些结构有助于揭示C-的位置和潜在作用。当蛋白质游离或与 DNA 结合时，这些知识将指导我们在实验室中设计新的实验，并引导我们更好地了解这个在植物防御病原体中发挥重要作用的蛋白质家族。