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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 铜菜（BRS）是类固醇激素，在植物中起着重要作用。细胞表面受体激酶BRI1感知到BR信号，该细胞表面受体激酶BRI1启动了一系列蛋白质磷酸化，从而导致核基因表达和细胞反应。下游BR信号传导涉及细胞质GSK3/shaggy样激酶BIN2。 BR灭活BIN2，它磷酸化了转录因子BZR1和BZR2/BES1。我们的蛋白质组学研究与遗传方法结合使用了几个缺失的联系，包括将信号从BRI1激酶传递到下游成分的BSK激酶。这项研究的目的是进一步了解生化和蛋白质组学水平的BR信号转导。首先，我们将使用串联亲和力纯化和质谱分析来鉴定BZR1相互作用的蛋白质。黄铜固醇通过诱导转录因子BZR1的去磷酸化而促进生长，但是脱磷酸酯BZR1仍然难以捉摸的磷酸化酶仍然无法使用。遗传和芯片芯片数据表明，BZR1可能与不同的转录因子相互作用以调节基因表达。这项研究将极大地有助于鉴定磷酸酶并阐明转录因子网络。其次，我们将确定该途径几个关键成分的磷酸化位点（BSK，BSUS，CDGS，BIN2）。第三，为了了解BR信号如何调节BIN2活性，我们将净化BIN2相互作用的蛋白质并使用MS分析识别它们。将使用遗传和转基因实验研究所鉴定蛋白的功能。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具有广泛的影响。