相关实验视频
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Robust 3D DNA FISH Using Directly Labeled Probes
Published on: August 15, 2013
通过多种细胞因子对DNA结合蛋白的氨酸酸化
A C Larner1, M David, G M Feldman
1Division of Cytokine Biology, Center for Biologics Evaluation and Research, Bethesda, MD 20892.
概括
像IL-3和GM-CSF这样的细胞因子通过铁酸化激活转录因子,但不会诱导FcRI基因表达. 干扰素- (IFN-) 和IL-10通过含有p91的复合体诱导FcRI的表达.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 细胞信号传递 细胞信号传递
背景情况:
- 干扰素-α (IFN-α) 和干扰素-gamma (IFN-gamma) 通过转录因子的氨酸酸化来调节基因表达,特别是干扰素刺激的基因因子-3 (ISGF-3) 的p91成分.
- 干扰素激活复合物与早期反应基因促进体中的增强剂结合,例如Fc玛受体I (Fc玛RI) 基因.
- 了解细胞因子介导的基因调节对于免疫反应调节至关重要.
研究的目的:
- 调查除了干扰素之外,其他细胞因子是否可以激活与IFN-马响应区域 (GRR) 结合的转录因子.
- 确定氨酸酸化和特定蛋白质成分在细胞因子诱导的基因激活中的作用.
- 检查各种细胞因子对诱导Fc玛RI基因表达的影响.
主要方法:
- 用特定的细胞因子治疗人类外周血液单细胞或基因细胞:IL-3,IL-5,IL-10,GM-CSF,IFN-gamma.
- 使用电泳运动转移试验,分析识别Fc马RI促进体GRR的DNA结合蛋白.
- 在激活的DNA结合复合体内通过西式涂抹或类似技术识别氨酸酸化蛋白质.
主要成果:
- 介质素-3 (IL-3),IL-5,IL-10和花细胞巨菌群刺激因子 (GM-CSF) 激活了识别Fc gamma RI GRR的DNA结合蛋白.
- 氨酸酸化对于这些GRR结合复合物的组装至关重要.
- 含有p91蛋白的IFN-和IL-10诱导复合体,导致FcRIRNA诱导.
- IL-3和GM-CSF激活复合物含有80kDa的氨酸酸化蛋白,但没有诱导Fc玛RIRNA.
- 转基因-CSF和IL-3预处理抑制了IFN-马诱导的Fc马RIRNA.
结论:
- 多种细胞因子,包括IL-3和GM-CSF,可以通过氨酸酸化激活转录因子,扩展到干扰素之外.
- 激活转录因子复合体 (p91与80kDa) 的特定蛋白质组成决定了下游基因表达结果.
- 细胞因子信号通路在调节基因诱导方面表现出特异性,Fc gamma RI表达的差异效应证明了这一点.
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