MOLECULAR MECHANISMS CYTOKINE RECEPTOR SIGNALING

细胞因子受体信号转导的分子机制

• 批准号: 6173091
• 负责人: Stephanie S Watowich
• 金额: $ 24.99万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173091
• 关键词: JAK kinase; biological signal transduction; cell differentiation; cell growth regulation; cell proliferation; chimeric proteins; complementary DNA; cytokine receptors; dimer; enzyme activity; erythroid stem cell; erythroleukemia; erythropoiesis; erythropoietin; growth factor receptors; immunoprecipitation; laboratory mouse; molecular site; mutant; protein binding; protein structure; receptor expression; tissue /cell culture; western blottings

DESCRIPTION: Cytokines act through specific membrane receptors to regulate hematopoietic cell development. A number of intracellular signaling pathways are stimulated by activated cytokine receptors; however, the molecular events involved in initiation of signal transduction, and how the different pathways control hematopoietic cell survival, proliferation, and differentiation, are largely unknown. Abnormal hematopoietic cell growth can lead to the development of disease, including myeloproliferative disorders, leukemia, or lymphoma. In addition, underproduction of crucial hematopoietic cell types can compromise health and survival. The overall goals of this project are to define the role specific protein:protein oligomerization events play in activation of cytokine receptor signal transduction, and to determine the specificity of Jak and Stat signals in erythroid development. In Aim 1 the role dimerization of the receptor cytoplasmic region plays in activation of downstream signal transduction pathways will be examined, using the erythropoietin receptor (EpoR) as a model of hemodimeric cytokine receptors. Specific dimerization motifs will be fused to the EpoR cytoplasmic tail. The effects of cytoplasmic tail dimerization, in the absence or presence of plasma membrane association, on the activation of signal transduction pathways leading to cell proliferation will be determined. In Aim 2 the role of homo-or heterodimerization of Jak protein tyrosine kinases in the role of regulation of intrinsic kinase activity will be studied. Chimeric EpoRs containing Jak1 or Jak3 binding domains will be generated and it will be determined if ligand-induced receptor dimerization activates the respective Jak kinase. Dimerization motifs will also be fused directly to Jak1, Jak2, or Jak3 cDNAs to determine if homo- or heterodimerization between specific Jaks regulates their activity. In the third aim, the investigator will determine if signals from Jak2 and Stat5 are specific for erythroid development by generating chimeric EpoRs with altered Jak or Stat signal transduction potential. These will be introduced into growth-factor dependent hematopoietic cells to analyze effects on cell proliferation in vitro. Constitutively active forms of the chimeric receptors will be introduced into hematopoietic progenitor cells and in vitro erythroid differentiation analyzed by colony-forming assays. Effects on in vivo erythropoiesis and contribution to development of leukemia will be analyzed in mice infected with recombinant retroviruses.

描述：细胞因子通过特定的膜受体发挥作用来调节 造血细胞发育。 多种细胞内信号传导 途径受到激活的细胞因子受体的刺激；然而， 参与信号转导启动的分子事件，以及如何 不同的途径控制造血细胞的存活、增殖和 的分化，很大程度上是未知的。 造血细胞生长异常 可能导致疾病的发展，包括骨髓增生性疾病 疾病、白血病或淋巴瘤。 此外，关键的生产不足 造血细胞类型会损害健康和生存。 整体 该项目的目标是定义特定蛋白质的作用：蛋白质 寡聚事件在细胞因子受体信号的激活中发挥作用 转导，并确定 Jak 和 Stat 信号的特异性 红细胞发育。 在目标 1 中，受体二聚化的作用 细胞质区域在下游信号转导的激活中发挥作用 将使用促红细胞生成素受体（EpoR）作为检查途径 血二聚体细胞因子受体模型。 特定的二聚化基序将 与 EpoR 细胞质尾融合。 细胞质尾的影响 二聚化，在不存在或存在质膜缔合的情况下， 信号转导途径的激活导致细胞增殖 将被确定。 在目标 2 中，Jak 同二聚体或异二聚体的作用 蛋白酪氨酸激酶在调节内在激酶中的作用 将研究活动。 含有 Jak1 或 Jak3 结合的嵌合 EpoR 将生成结构域并确定是否是配体诱导的 受体二聚化激活各自的 Jak 激酶。 二聚化 基序也将直接融合到 Jak1、Jak2 或 Jak3 cDNA 以确定 如果特定 Jaks 之间的同二聚或异二聚化调节其 活动。 在第三个目标中，研究人员将确定信号是否来自 Jak2 和 Stat5 通过生成嵌合体专门针对红细胞发育 EpoR 具有改变的 Jak 或 Stat 信号转导潜力。 这些将是 引入生长因子依赖性造血细胞进行分析 对体外细胞增殖的影响。 的本构活跃形式 嵌合受体将被引入造血祖细胞 以及通过集落形成试验分析体外红细胞分化。 对体内红细胞生成的影响及其对发育的贡献 将在感染重组逆转录病毒的小鼠中对白血病进行分析。

项目成果

Activation of erythropoietin signaling by receptor dimerization.

通过受体二聚化激活促红细胞生成素信号传导。

• DOI: 10.1016/s1357-2725(99)00075-8
• 发表时间: 1999
• 期刊: The international journal of biochemistry & cell biology
• 作者: Watowich,SS

Hematopoietic cell survival signals are elicited through non-tyrosine-containing sequences in the membrane-proximal region of the erythropoietin receptor (EPOR) by a Stat5-dependent pathway.

造血细胞存活信号是通过 Stat5 依赖性途径通过促红细胞生成素受体 (EPOR) 近膜区域中不含酪氨酸的序列引发的。

• DOI: 10.1016/j.exphem.2003.08.009
• 发表时间: 2003
• 期刊: Experimental hematology
• 影响因子: 2.6
• 作者: Yoon,Donghoon;Watowich,StephanieS
• 通讯作者: Watowich,StephanieS