晶体结构和循环二甲胺合成cGAS/DncV类核样转移酶的功能影响
Chia-Shin Yang1, Tzu-Ping Ko2, Chao-Jung Chen3,4
1Genomics BioSci & Tech Co. Ltd., New Taipei, 221, Taiwan.
Nature communications
|August 21, 2023
概括
研究人员阐明了循环二胺的合成,这对于细菌抗菌菌体防御至关重要. 他们确定了关键的蛋白质动机和通路,揭示了这些分子是如何产生的.
科学领域:
- 分子生物学分子生物学
- 生物化学 生化学
- 结构生物学 结构生物学
背景情况:
- 核酸第二信使调节重要的细胞过程.
- 循环二胺素参与细菌的抗菌体防御,但它们的合成尚不清楚.
- 循环核酸合成酶,cGAS/DncV类核酸转移酶 (CD-NTases),对于产生这些信使至关重要.
研究的目的:
- 为了确定循环二甲胺合成CD-NTases (CdnE) 的高分辨率结构.
- 阐明皮里米丁特异性的机制和循环二-皮里米丁的合成途径.
- 识别涉及基质识别和产品形成的关键残留物和基因.
主要方法:
- 采用X射线晶体学,以获得CdnE在apo,基板和中间结合状态中的高分辨率结构.
- 位点定向突变发生,以调查保存残留物在核酸特异性中的作用.
- 生物化学试验分析酶活性和产品形成.
主要成果:
- 高分辨率结构揭示了CdnE的apo,基板和中间结合状态.
- 一个保存的 (R/Q) xW 动机被确定为对胺素供体核酸特异性的关键.
- 在 (R/Q) xW 基因中发生的突变改变了对纯素核酸的特异性,产生混合的纯素-皮里米丁循环二核酸 (CDN).
- 乌拉在CdnE对细胞因子的优先结合解释了它对循环二-UMP (cUU) 的特异性.
- 通过pppU[3'-5']pU中间体,从中间体结合结构中推断出了cUU的一种新型合成途径,涉及2'3'-二结合,通过ppU[3'-5']pU中间体.
结论:
- 这项研究为CD-NTase活性中的pyrimidine选择提供了一个结构和机制框架.
- 在CD-NTases的C端环中保存的残留物对于确定酶特异性至关重要.
- 了解这些机制为操纵生物技术应用的CDN合成打开了道路.
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