SIGNAL TRANSDUCTION PATHWAYS IN HEMATOPOIESIS
造血中的信号转导途径
基本信息
- 批准号:6638531
- 负责人:
- 金额:$ 31.89万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1999
- 资助国家:美国
- 起止时间:1999-05-10 至 2004-04-30
- 项目状态:已结题
- 来源:
- 关键词:Drosophilidae JAK kinase biological signal transduction blood cells cell proliferation cell transformation erythropoietin genetic transcription genetically modified animals growth factor receptors guanosine triphosphate hematopoiesis hematopoietic growth factor molecular cloning mutant neoplastic transformation nuclear factor kappa beta phenotype phosphorylation protein structure function protein tyrosine kinase receptor expression suppressor mutations transcription factor transfection
项目摘要
We postulate that there is extensive crosstalk between hematopoietic signal transduction pathways. We propose that this crosstalk helps to regulate the activation and termination of signals emanating from the surface of activated hematopoietic cells. To begin to address this hypothesis, two constitutively active receptors (cEpoR and CD8-cEyk), known to activate the JAK/STAT pathway, were introduced into murine hematopoietic (Ba/F3) cells. Both caused factor independence and transformation, as a result of activation of the JAK/STAT and Ras/Raf pathways. However, signaling events initiated by cEpoR and CD8-cEyk were clearly different. To further evaluate these differences, we have developed an in vivo assay in Drosophila blood cells. We take advantage of the fact that signaling pathways are remarkably conserved among metazoans. cDNAs for both receptors have been placed under the control of Gal4 transcriptional regulatory elements, and injected into Drosophila embryos. Transgenic flies were isolated and crossed with Drosophila lines expressing Gal4 protein under the control of a heat shock promoter. Upon heat shock, expression of the vertebrate receptors is induced, resulting in leukemia-like proliferation of blood cells and formation of melanotic tumors. This offers a powerful, novel assay to probe signal transduction circuits in vivo. We will further refine this assay, and determine the participation of the JAK/STAT, Ras/Raf, NF-kappaB and JNK pathways in hemocyte proliferation and tumorigenesis in genetic experiments. We will test whether loss of function mutants in these pathways can suppress the melanotic tumor phenotype. In addition, we will use this system to characterize a hyperphosphorylated STAT5 mutant that we have recently generated. We have shown that this STAT5 mutant is resistant to dephosphorylation. We expect to find that it confers a gain of function phenotype. Finally, we will exploit a genetic screen in Drosophila to isolate novel genes that interact with known signal transduction pathways to suppress the melanotic tumor phenotype. Viable and fertile flies, which all show melanotic tumors, will be crossed with mutant stocks carrying chromosomal aberrations, P-element insertions, or chemically- induced point mutations. Suppression of the melanotic tumor phenotype in the F1 generation will serve as the criterion to identify accessory factors that modulate known hematopoietic signal transduction pathways. This series of experiments should produce new insights into hematopoietic signal transduction that could not be obtained from standard biochemical experiments, or from gene manipulations in mice.
我们假设造血信号转导途径之间存在广泛的串扰。 我们认为这种串扰有助于调节从活化的造血细胞表面发出的信号的激活和终止。 为了开始解决这一假设,将已知可激活 JAK/STAT 途径的两种组成型活性受体(cEpoR 和 CD8-cEyk)引入小鼠造血 (Ba/F3) 细胞中。 由于 JAK/STAT 和 Ras/Raf 途径的激活,两者都会引起因子独立和转化。 然而,cEpoR 和 CD8-cEyk 引发的信号传导事件明显不同。 为了进一步评估这些差异,我们开发了果蝇血细胞的体内测定法。 我们利用了信号通路在后生动物中非常保守的事实。 两种受体的 cDNA 已置于 Gal4 转录调控元件的控制下,并注射到果蝇胚胎中。 分离转基因果蝇并与在热休克启动子控制下表达Gal4蛋白的果蝇系杂交。 热休克后,脊椎动物受体的表达被诱导,导致血细胞的白血病样增殖和黑色素瘤的形成。这提供了一种强大的、新颖的检测方法来探测体内信号转导电路。 我们将进一步完善该测定,并在基因实验中确定JAK/STAT、Ras/Raf、NF-kappaB和JNK通路在血细胞增殖和肿瘤发生中的参与。 我们将测试这些途径中功能缺失的突变体是否可以抑制黑色素肿瘤表型。 此外,我们将使用该系统来表征我们最近生成的过度磷酸化 STAT5 突变体。 我们已经证明该 STAT5 突变体能够抵抗去磷酸化。 我们期望发现它赋予功能表型的增益。 最后,我们将利用果蝇中的遗传筛选来分离与已知信号转导途径相互作用的新基因,以抑制黑色素瘤表型。存活且可育的果蝇均表现出黑色素瘤,将与携带染色体畸变、P 元件插入或化学诱导点突变的突变体杂交。 F1代黑色素肿瘤表型的抑制将作为识别调节已知造血信号转导途径的辅助因子的标准。 这一系列的实验应该会对造血信号转导产生新的见解,而这是从标准生化实验或小鼠基因操作中无法获得的。
项目成果
期刊论文数量(6)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Differential requirement for STAT by gain-of-function and wild-type receptor tyrosine kinase Torso in Drosophila.
果蝇功能获得和野生型受体酪氨酸激酶躯干对 STAT 的不同需求。
- DOI:10.1242/dev.129.18.4241
- 发表时间:2002
- 期刊:
- 影响因子:0
- 作者:Li,WillisX;Agaisse,Herve;Mathey-Prevot,Bernard;Perrimon,Norbert
- 通讯作者:Perrimon,Norbert
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BERNARD MATHEY-PREVOT其他文献
BERNARD MATHEY-PREVOT的其他文献
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{{ truncateString('BERNARD MATHEY-PREVOT', 18)}}的其他基金
DERIVATION OF MODEL SYSTEMS FOR HEMATOPOIETIC GROWTH FACTOR RECEPTOR FUNCTION
造血生长因子受体功能模型系统的推导
- 批准号:
6105669 - 财政年份:1998
- 资助金额:
$ 31.89万 - 项目类别:
DERIVATION OF MODEL SYSTEMS FOR HEMATOPOIETIC GROWTH FACTOR RECEPTOR FUNCTION
造血生长因子受体功能模型系统的推导
- 批准号:
6239205 - 财政年份:1997
- 资助金额:
$ 31.89万 - 项目类别:
GENE REGULATION OF THE CYTOKINES IL-3 AND GM-CSF
细胞因子 IL-3 和 GM-CSF 的基因调控
- 批准号:
3242635 - 财政年份:1990
- 资助金额:
$ 31.89万 - 项目类别:
GENE REGULATION OF THE CYTOKINES IL-3 AND GM-CSF
细胞因子 IL-3 和 GM-CSF 的基因调控
- 批准号:
3242636 - 财政年份:1990
- 资助金额:
$ 31.89万 - 项目类别:
GENE REGULATION OF THE CYTOKINES IL3 AND GM CSF
细胞因子 IL3 和 GM CSF 的基因调控
- 批准号:
2141906 - 财政年份:1990
- 资助金额:
$ 31.89万 - 项目类别:
GENE REGULATION OF THE CYTOKINES IL-3 AND GM-CSF
细胞因子 IL-3 和 GM-CSF 的基因调控
- 批准号:
3242634 - 财政年份:1990
- 资助金额:
$ 31.89万 - 项目类别: