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: 8169732
• 负责人: ZHIYONG WANG
• 金额: $ 4.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169732
• 关键词: Affinity Chromatography; Biochemical; Cell Surface Receptors; Computer Retrieval of Information on Scientific Projects Database; DNA Binding; Funding; Gene Expression; Genetic; Goals; Grant; Institution; Mass Spectrum Analysis; Molecular; Nuclear; Phosphorylation; Phosphorylation Site; Phosphotransferases; Plants; Proteins; Proteomics; Research; Research Personnel; Research Project Grants; Resources; Role; Signal Transduction; Site-Directed Mutagenesis; Source; Steroids; Testing; Transgenic Organisms; United States National Institutes of Health; multicatalytic endopeptidase complex; 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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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. BIN2 phosphorylates the transcription factors BZR1 and BZR2/BES1, and this phosphorylation inhibits their DNA binding and nuclear localization and promotes their degradation by the proteasome. The goal of this study is to understand BR signal transduction at the biochemical and proteomic levels. First, we will identify the BIN2-phosphorylation sites of BZR1 using mass spectrometry (MS). The importance of some of the phosphorylation sites will be further tested by site-directed-mutagenesis and transgenic experiments. Second, we will identify the BZR1-interacting proteins using tandem affinity purification and mass spectrometry analysis. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 油菜素甾醇（BR）是在植物中具有重要作用的类固醇激素。 BR 信号由细胞表面受体激酶 BRI1 感知，启动一系列蛋白磷酸化，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。 BIN2 磷酸化转录因子 BZR1 和 BZR2/BES1，这种磷酸化抑制它们的 DNA 结合和核定位，并促进它们被蛋白酶体降解。本研究的目的是在生化和蛋白质组水平上了解 BR 信号转导。首先，我们将使用质谱 (MS) 鉴定 BZR1 的 BIN2 磷酸化位点。一些磷酸化位点的重要性将通过定点诱变和转基因实验进一步测试。其次，我们将使用串联亲和纯化和质谱分析来鉴定 BZR1 相互作用蛋白。第三，为了了解 BR 信号传导如何调节 BIN2 活性，我们将纯化 BIN2 相互作用蛋白并使用 MS 分析对其进行鉴定。将使用遗传和转基因实验来研究已鉴定蛋白质的功能。该研究项目将增进我们对植物类固醇反应分子机制的理解，这将对我们对类固醇作用的总体理解产生广泛的影响。