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一个类似菌体核的隔间保护DNA免受CRISPR核酶的影响
Senén D Mendoza1, Eliza S Nieweglowska2, Sutharsan Govindarajan1,3
1Department of Microbiology and Immunology, University of California San Francisco, San Francisco, CA, USA.
Nature
|December 11, 2019
概括
大菌体 ΦKZ 通过构建一个类似于核的隔间来逃避细菌免疫系统,保护其DNA免受核酶的影响. 这种蛋白质屏障保护了菌体基因组,使得宿主体内的有效复制成为可能.
科学领域:
- 微生物学
- 病毒学
- 分子生物学
背景情况:
- 病毒,包括菌体 (菌体),必须克服宿主免疫防御以复制.
- 像CRISPR-Cas和限制修饰这样的细菌免疫系统是针对外来核酸的.
- 大型菌体拥有大型基因组和复杂的复制策略,需要强大的逃避机制.
研究的目的:
- 调查大菌体对细菌防御系统使用的免疫逃避策略.
- 确定 ΦKZ 在感染期间如何保护其DNA免受宿主编码的核酶的影响.
- 通过隔间操纵来探索菌体易感性的可能性.
主要方法:
- 使用Pseudomonas aeruginosa和大菌体 ΦKZ进行感染测定.
- 菌体诱导的蛋白质区的特征.
- 测试 ΦKZ 对各种CRISPR-Cas酶 (Cas3,Cas9,Cas12a,Cas13a) 和限制酶 (HsdRMS,EcoRI) 的耐药性.
- 在宿主细胞内重新定位限制酶.
主要成果:
- 大菌体 ΦKZ 构建了一个类似于核的蛋白质区,将其DNA从宿主免疫核酶中隔离.
- 对于多个向DNA的CRISPR- Cas和限制酶来说, ΦKZ DNA 仍然是不可访问的.
- 将EcoRI重新定位到隔间使 ΦKZ易受DNA向.
- ΦKZ对向RNA的CRISPR- Cas酶Cas13a敏感,这表明菌体mRNA的细胞质局部化.
结论:
- 伪虫巨型菌体利用一种蛋白质的区间来保护它们的基因组免受广泛的针对DNA的细菌免疫系统的影响.
- 这种分离是免疫宿主中高效菌体复制的关键策略.
- 菌体对RNA向系统的敏感性凸显了mRNA局部化在病毒感染动态中的重要性.
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