人类cGAS二核酸结合特异性的结构引导重编程
Philip J Kranzusch1, Amy S Y Lee2, Stephen C Wilson3
1Department of Molecular & Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Howard Hughes Medical Institute (HHMI), University of California, Berkeley, Berkeley, CA 94720, USA.
Cell
|August 19, 2014
概括
这项研究揭示了细菌酶和人类cGAS如何合成具有特定联系的循环二核酸 (CDN). 了解这种机制可以澄清细菌病原和天生的免疫路径.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
背景情况:
- 循环二核酸 (CDN) 是细菌病原和哺乳动物天生的免疫中至关重要的信号分子.
- 酶cGAS产生2'-5'cGAMP,对免疫刺激至关重要,但其链接特异性尚不清楚.
- 具有与cGAS相似功能的 Prokaryotic 酶在很大程度上没有表征.
研究的目的:
- 阐明在cGAS介导的循环二核酸合成中的2'-5'二结合特异性的基础分子机制.
- 为了研究细菌酶DncV的结构和功能,作为cGAS的细胞原生同类物.
- 建立细菌信号通路和哺乳动物天生的免疫之间的机械同质性.
主要方法:
- 霍乱病毒DncV和人类cGAS的高分辨率晶体结构.
- 生物化学试验分析CDN合成和链接特异性.
- 部位定向突变发生,以重新编程人类cGAS活性部位和细胞分析,以测试STING激活.
主要成果:
- 作为一种 prokaryotic 酶的 DncV 在循环二核酸合成中与人类的 cGAS 有机相似之处.
- 晶体结构显示,DncV和cGAS通过以相反方向进行的顺序反应合成CDN.
- 重编程人类的cGAS活性部位使得3'-5'cGAMP的产生成为可能,选择性地激活了替代性STING适应基因.
结论:
- cGAS和DncV的活性位点配置决定了循环二核酸的特定基结化学 (2'-5'或3'-5') .
- 这种联系特异性控制了细菌和哺乳动物系统的下游信号通路.
- 细菌病变因子和先天性免疫酶之间的机械同源性为保存的信号机制提供了洞察力.
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