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: 7724170
• 负责人: ZHIYONG WANG
• 金额: $ 1.29万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7724170
• 关键词: Affinity Chromatography; Biochemical; Cell Surface Receptors; Cell membrane; Computer Retrieval of Information on Scientific Projects Database; DNA Binding; Funding; Gene Expression; Genetic; Goals; Grant; Institution; Mass Spectrum Analysis; Metals; Methods; Molecular; Nuclear; Phosphoproteins; 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; Two-Dimensional Gel Electrophoresis; United States National Institutes of Health; multicatalytic endopeptidase complex; research study; response; steroid hormone; transcription factor; two-dimensional

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. Phosphorylation of the transcription factors 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, in order to understand how BR signaling regulates BIN2 activity, we will purify BIN2-interacting proteins and identify them using MS analysis. Third, we are identifying BR-regulated proteins using two-dimensional difference gel electrophoresis (2-D DIGE) and iTRAQ mass spectrometry. We will identify BR-regulated proteins by performing proteomic analysis of total protein, plasma membrane, and phosphoprotein fractions. Forth, we plan to study BR regulated protein phosphorylation using immobilized metal affinity chromatography and the iTRAQ method. 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.

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