PROTEOMIC STUDY OF BRASSINOSTEROID SIGNAL TRANSDUCTION

油菜素甾醇信号转导的蛋白质组学研究

• 批准号: 8363740
• 负责人: ZHIYONG WANG
• 金额: $ 1.44万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363740
• 关键词: Affinity Chromatography; Biochemical; Cell Surface Receptors; Data; Funding; Gene Expression; Genetic; Goals; Grant; Growth; Link; Mass Spectrum Analysis; Molecular; National Center for Research Resources; Nuclear; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Plants; Principal Investigator; Protein Dephosphorylation; Proteins; Proteomics; Research; Research Infrastructure; Research Project Grants; Resources; Role; Signal Transduction; Source; Steroids; Transgenic Organisms; United States National Institutes of Health; cost; research study; response; steroid hormone; transcription factor

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. Brassinosteriods (BRs) are steroid hormones with important roles in plants. BR signal is perceived by the cell-surface receptor kinase BRI1, which initiates a cascade of protein phosphorylation leading to nuclear gene expression and cellular responses. Downstream BR signaling involves the cytoplasmic GSK3/SHAGGY-like kinase BIN2. BR inactivate BIN2, which phosphorylates the transcription factors BZR1 and BZR2/BES1. Our proteomic studies combined with genetic approaches have identified several missing links, including BSKs kinases that transduce signals from BRI1 kinases to downstream components. The goal of this study is to further understand BR signal transduction at the biochemical and proteomic levels. First,we will identify the BZR1-interacting proteins using tandem affinity purification and mass spectrometry analysis. Brassinosteroid promotes growth by inducing dephosphorylation of transcription factor BZR1, but the phosphatase(s) that desphosphorylate BZR1 still remain(s) elusive. Genetic and CHIP-CHIP data have suggested BZR1 might interact with different transcription factors to regulate gene expression. This study will greatly help to identify the phosphotase and to elucidate the networks of transcription factors. Second, we will identify phosphorylation sites of several key components of this pathway (BSK, BSUs, CDGs, BIN2). Third, in order to understand how BR signaling regulates BIN2 activity, we will purify BIN2-interacting proteins and identify them using MS analysis. The functions of the identified proteins will be studied using genetic and transgenic experiments. This research project will advance our understanding of the molecular mechanism for steroid responses in plants, which will have broad implications in our understanding of steroid actions in general.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 甾体类化合物是一类在植物中具有重要作用的甾体激素。BR信号由细胞表面受体激酶BRI 1感知，其启动蛋白磷酸化级联反应，导致核基因表达和细胞反应。下游BR信号传导涉及细胞质GSK 3/SHAGGY样激酶BIN 2。BR β BIN 2，磷酸化转录因子BZR 1和BZR 2/BES 1。我们的蛋白质组学研究结合遗传学方法已经确定了几个缺失的环节，包括BSKs激酶，它将信号从BRI 1激酶传递到下游组分。本研究的目标是在生化和蛋白质组水平上进一步了解BR信号传导。首先，我们将使用串联亲和纯化和质谱分析来鉴定BZR 1相互作用蛋白。类维生素C通过诱导转录因子BZR 1的去磷酸化来促进生长，但是使BZR 1去磷酸化的磷酸酶仍然是难以捉摸的。遗传和CHIP-CHIP数据表明BZR 1可能与不同的转录因子相互作用以调节基因表达。本研究将有助于磷酸酶的鉴定和转录因子网络的阐明。其次，我们将确定该途径的几个关键组分（BSK，BSU，CDG，BIN 2）的磷酸化位点。第三，为了了解BR信号如何调节BIN 2活性，我们将纯化BIN 2相互作用蛋白，并使用MS分析鉴定它们。将使用遗传和转基因实验来研究所鉴定的蛋白质的功能。该研究项目将促进我们对植物中类固醇反应的分子机制的理解，这将对我们对类固醇作用的理解产生广泛的影响。