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大肠杆菌CRISPR-Cas系统对具有不同生活方式和发育策略的裂解性噬菌体的作用。

The action of Escherichia coli CRISPR-Cas system on lytic bacteriophages with different lifestyles and development strategies.

作者信息

Strotskaya Alexandra, Savitskaya Ekaterina, Metlitskaya Anastasia, Morozova Natalia, Datsenko Kirill A, Semenova Ekaterina, Severinov Konstantin

机构信息

Skolkovo Institute of Science and Technology, Moscow, Russia.

Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia.

出版信息

Nucleic Acids Res. 2017 Feb 28;45(4):1946-1957. doi: 10.1093/nar/gkx042.

Abstract

CRISPR-Cas systems provide prokaryotes with adaptive defense against bacteriophage infections. Given an enormous variety of strategies used by phages to overcome their hosts, one can expect that the efficiency of protective action of CRISPR-Cas systems against different viruses should vary. Here, we created a collection of Escherichia coli strains with type I-E CRISPR-Cas system targeting various positions in the genomes of bacteriophages λ, T5, T7, T4 and R1-37 and investigated the ability of these strains to resist the infection and acquire additional CRISPR spacers from the infecting phage. We find that the efficiency of CRISPR-Cas targeting by the host is determined by phage life style, the positions of the targeted protospacer within the genome, and the state of phage DNA. The results also suggest that during infection by lytic phages that are susceptible to CRISPR interference, CRISPR-Cas does not act as a true immunity system that saves the infected cell but rather enforces an abortive infection pathway leading to infected cell death with no phage progeny release.

摘要

CRISPR-Cas系统为原核生物提供了抵御噬菌体感染的适应性防御机制。鉴于噬菌体用于克服宿主的策略多种多样,人们可以预期CRISPR-Cas系统对不同病毒的保护作用效率会有所不同。在这里,我们构建了一组带有I-E型CRISPR-Cas系统的大肠杆菌菌株,该系统靶向噬菌体λ、T5、T7、T4和R1-37基因组中的不同位置,并研究了这些菌株抵抗感染以及从感染噬菌体中获取额外CRISPR间隔序列的能力。我们发现,宿主对CRISPR-Cas的靶向效率由噬菌体生活方式、基因组中靶向原间隔序列的位置以及噬菌体DNA的状态决定。结果还表明,在受CRISPR干扰影响的烈性噬菌体感染期间,CRISPR-Cas并非作为一种真正的免疫机制拯救被感染细胞,而是强制导致一种流产感染途径,致使被感染细胞死亡且无噬菌体后代释放。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8437/5389539/2b6c2cded2df/gkx042fig1.jpg

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