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RAG1调解信号序列识别和RAG2在V(D) J重组中的招募
M J Difilippantonio1, C J McMahan, Q M Eastman
1Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
Cell
|October 18, 1996
概括
单独RAG1蛋白在V(D) J重组过程中识别DNA重组信号,与RAG2一起.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- V(D) J重组是适应性免疫的一个关键过程,涉及由RAG1和RAG2蛋白调解的DNA裂变.
- 研究RAG1和RAG2的体外DNA结合相互作用一直是具有挑战性的.
- 了解这些相互作用是解读适应性免疫系统发育机制的关键.
研究的目的:
- 独立于RAG2.2,研究RAG1的DNA结合能力.
- 为了识别由RAG1.1.识别的特定DNA元素.
- 阐明RAG2在信号定位中的作用及其对RAG1.1的依赖性.
主要方法:
- 采用了体内一种混合基因DNA结合试验.
- 在重组信号中研究了RAG1与非胺和胺序列的结合.
- 分析了间隔器长度 (12bp vs. 23bp) 对RAG2局部化的影响.
主要成果:
- 只有RAG1才能识别重组信号,主要是通过非amer序列.
- 七度体序列增强了RAG1的结合亲和力.
- 对细菌逆转酶同类的RAG1区域对于DNA结合至关重要.
- 对信号的RAG2定位是RAG1依赖的,并且在12bp间距时更有效.
结论:
- RAG1 具有对V(D) J重组信号的内在DNA结合能力.
- RAG1与非聚体的相互作用是基本的,而六聚体提供了增强.
- RAG2的招募由RAG1促进,并受到信号结构的影响,这表明了协调的绑定机制.
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