TIF AND ERYTHROLEUKEMIA CELL DIFFERENTIATION

TIF 和红白血病细胞分化

• 批准号: 2100592
• 负责人: WEI DAI
• 金额: $ 21.17万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2100592
• 关键词: affinity chromatography; biological signal transduction; cell cycle; cell differentiation; colony stimulating factor; enzyme activity; erythroid stem cell; erythroleukemia; erythropoietin; gene expression; growth factor receptors; hematopoiesis; immunoprecipitation; intermolecular interaction; neoplastic cell; phenotype; protein tyrosine kinase; receptor expression; tissue /cell culture; yeasts; affinity chromatography; biological signal transduction; cell cycle; cell differentiation; colony stimulating factor; enzyme activity; erythroid stem cell; erythroleukemia; erythropoietin; gene expression; growth factor receptors; hematopoiesis; immunoprecipitation; intermolecular interaction; neoplastic cell; phenotype; protein tyrosine kinase; receptor expression; tissue /cell culture; yeasts

DESCRIPTION: This revised application describes studies to elucidate the functional role and molecular mechanism of action of tif, a new member of the axl family of receptors. Tif, which was cloned by RTPCR from a K562 library, is an extremely interesting receptor in that it bears extracellular fibronectin domains as well as Ig domains, suggesting a role in cell-matrix attachment and signalling. In addition, its pattern of RNA expression, gonad>brain>other tissues, suggests a potential role in primitive cell maintenance and self-renewal. In this application, the Applicant proposes to: 1) Study the transforming potential of tif by enforced over- expression; 2) Investigate the role of tif in hematopoietic determination and differentiation by analyzing the phenotype of both K562 and Ba/F3 cells constitutively expressing activated tif, and by differentiating ES cells with a targeted gene disruption of the tif locus; 3)Identify the immediate downstream components of the tif signaling pathway via immonocoprecipitation and via the yeast two-hybrid system; 4)Search for a functional interaction between tif and the Epo receptor by enforced expression of an EpoR/tif chimera, analyzing signal transduction and cellular phenotype in transfected Ba/F3 and K562 cells; and 5)Identify and clone the tif ligand via expression or affinity column chromatography. These studies are designed to provide the first important steps towards understanding the role of Tif in hematopoietic differentiation.

描述：此修订的应用程序描述了阐明的研究 TIF的功能作用和分子机理，一种新的 AXL受体家族的成员。 TIF，由RTPCR克隆 来自K562库，是一个非常有趣的受体 它带有细胞外纤连蛋白结构域以及IG 域，表明在细胞矩阵附着和信号传导中起作用。 此外，它的RNA表达模式，性腺>大脑>其他 组织表明在原始细胞维持中具有潜在的作用 和自我更新。 在此应用程序中，申请人建议： 1）研究强制性过度的TIF的转换潜力 表达; 2）研究TIF在造血的作用 通过分析表型来确定和分化 K562和BA/F3细胞均可表达活化的TIF， 并通过靶向基因破坏区分ES细胞 TIF基因座； 3）确定立即的下游组件 TIF信号通路通过免疫药物和酵母 两个杂交系统； 4）搜索TIF之间的功能相互作用 和通过Epor/TIF强制表达的EPO受体 嵌合体，分析信号转导和细胞表型 转染的BA/F3和K562细胞； 5）识别和克隆TIF 配体通过表达或亲和力色谱柱色谱法。 这些 研究旨在提供第一个重要步骤 了解TIF在造血分化中的作用。