Mechanism of Lnk function in cytokine receptor signaling

Lnk在细胞因子受体信号传导中的作用机制

基本信息

批准号：
7640868
负责人：
Wei Tong
金额：
$ 15.92万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7640868
关键词：
Adaptor Signaling Protein Address Affect Binding Biochemical Biology Blood Platelets Bone Marrow Cell Culture System Cells Complex Cytokine Receptors Defect Development Dimerization Docking Down-Regulation Erythrocytes Erythroid Erythropoiesis Erythropoietin Family member Functional disorder Genes Genetic Grant Hematopoiesis Hematopoietic Homo Immunoprecipitation JAK2 gene Janus kinase Knowledge Libraries Light Lymphocyte Maps Mass Spectrum Analysis Mediating Megakaryocytes Minor Modeling Mus Oncogenic PH Domain Peptide Library Phosphopeptides Phosphorylation Phosphotyrosine Physiological Play Predisposition Production Proline Protein Tyrosine Kinase Proteins Proteomics Receptor Inhibition Receptor Signaling Recombinants Regulation Research Research Personnel Role Screening procedure Signal Pathway Signal Transduction Signal Transduction Pathway Structure System Testing Thrombopoietin Training Tyrosine cytokine homologous recombination in vivo leukemia leukemogenesis mutant novel progenitor programs receptor recombinant peptide response skills src Homology Region 2 Domain three dimensional structure two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Erythropoietin (Epo) and thrombopoietin (Tpo) are the primary cytokines regulating red blood cell production, and megakaryocyte and platelet development, respectively. The adaptor protein Lnk negatively regulates signaling transduction of the Epo receptor (EpoR) and Tpo receptor (mpl), thereby downregulating Tpo-mediated megakaryocytopoiesis and Epo-mediated erythropoiesis. Lnk inhibits the activity of cytokine receptor integral partner, the JAK2 tyrosine kinase. Furthermore, the Lnk SH2 domain is essential for its inhibitory functions, and the functional interaction between Lnk and the receptor/JAK2 complex is phosphorylation-dependent. The purpose of this grant is to unravel the mechanism that encodes how Lnk controls the quantitative response to cytokines, and whose dysregulation predisposes to leukemia. To accomplish this, biochemical, cellular, and mouse genetic approaches will be used with the focus on the Lnk SH2 domain and tyrosine residues. Recombinant peptide library screens will establish Lnk SH2 domain binding motifs and mass spectrometry (MS) will identify Lnk SH2 domain interacting proteins in vivo. Novel primary hematopoietic cell culture systems will be developed to investigate the interactions among Lnk, cytokine receptors, and JAK2. Moreover, the Lnk tyrosines that are phosphorylated in vivo and important for its function will be dissected. Importantly, the mechanisms of Lnk regulatory function will be tested within physiological contexts using gene-deficient primary hematopoietic cells and generating gene-ablated mice. Unchecked and sustained cytokine receptor signaling leads to oncogenic transformation, therefore understanding the cellular mechanisms by which Lnk controls the amplitude and duration of receptor signaling will shed significant light on cytokine receptor regulation in general. Through this proposed research, the applicant will receive training in structural and proteomics biology and will gain the necessary skills and knowledge to become an independent investigator.

描述（由申请人提供）：促红细胞生成素（Epo）和血小板生成素（Tpo）分别是调节红细胞生成以及巨核细胞和血小板发育的主要细胞因子。接头蛋白 Lnk 负向调节 Epo 受体 (EpoR) 和 Tpo 受体 (mpl) 的信号转导，从而下调 Tpo 介导的巨核细胞生成和 Epo 介导的红细胞生成。 Lnk 抑制细胞因子受体整合伙伴 JAK2 酪氨酸激酶的活性。此外，Lnk SH2 结构域对其抑制功能至关重要，并且 Lnk 与受体/JAK2 复合物之间的功能相互作用是磷酸化依赖性的。这笔资助的目的是揭示 Lnk 如何控制对细胞因子的定量反应的编码机制，以及其失调容易导致白血病。为了实现这一目标，将使用生化、细胞和小鼠遗传方法，重点关注 Lnk SH2 结构域和酪氨酸残基。重组肽库筛选将建立 Lnk SH2 结构域结合基序，质谱 (MS) 将鉴定体内 Lnk SH2 结构域相互作用蛋白。将开发新型原代造血细胞培养系统来研究 Lnk、细胞因子受体和 JAK2 之间的相互作用。此外，还将解剖体内磷酸化且对其功能很重要的 Lnk 酪氨酸。重要的是，Lnk 调节功能的机制将在生理背景下使用基因缺陷的原代造血细胞和生成基因消除的小鼠进行测试。不受控制和持续的细胞因子受体信号传导会导致致癌转化，因此了解 Lnk 控制受体信号传导的幅度和持续时间的细胞机制将为细胞因子受体的一般调节提供重要的线索。通过这项拟议的研究，申请人将接受结构和蛋白质组生物学的培训，并将获得成为独立研究者所需的技能和知识。