CRISPR-Cas3 和 I 型限制修饰系统联手对抗肺炎克雷伯菌中 bla-IncF 质粒的转移。

CRISPR-Cas3 and type I restriction-modification team up against bla-IncF plasmid transfer in Klebsiella pneumoniae.

机构信息

Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai, China.

Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, Shanghai, China.

出版信息

BMC Microbiol. 2024 Jul 3;24(1):240. doi: 10.1186/s12866-024-03381-7.

DOI:10.1186/s12866-024-03381-7

PMID:38961341

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11223367/

Abstract

OBJECTIVE

We explored whether the Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification (R-M) systems are compatible and act together to resist plasmid attacks.

METHODS

932 global whole-genome sequences from GenBank, and 459 K. pneumoniae isolates from six provinces of China, were collected to investigate the co-distribution of CRISPR-Cas, R-M systems, and bla plasmid. Conjugation and transformation assays were applied to explore the anti-plasmid function of CRISPR and R-M systems.

RESULTS

We found a significant inverse correlation between the presence of CRISPR and R-M systems and bla plasmids in K. pneumoniae, especially when both systems cohabited in one host. The multiple matched recognition sequences of both systems in bla-IncF plasmids (97%) revealed that they were good targets for both systems. Furthermore, the results of conjugation assay demonstrated that CRISPR-Cas and R-M systems in K. pneumoniae could effectively hinder bla plasmid invasion. Notably, CRISPR-Cas and R-M worked together to confer a 4-log reduction in the acquisition of bla plasmid in conjugative events, exhibiting robust synergistic anti-plasmid immunity.

CONCLUSIONS

Our results indicate the synergistic role of CRISPR and R-M in regulating horizontal gene transfer in K. pneumoniae and rationalize the development of antimicrobial strategies that capitalize on the immunocompromised status of KPC-KP.

摘要

目的

我们探讨了簇状规律间隔短回文重复（CRISPR）-Cas 和限制修饰（R-M）系统是否兼容并共同作用以抵抗质粒攻击。

方法

从 GenBank 中收集了 932 个全球全基因组序列和来自中国六个省份的 459 株肺炎克雷伯菌分离株，以调查 CRISPR-Cas、R-M 系统和 bla 质粒的共分布。应用接合和转化试验来探索 CRISPR 和 R-M 系统的抗质粒功能。

结果

我们发现肺炎克雷伯菌中 CRISPR 和 R-M 系统与 bla 质粒的存在呈显著负相关，尤其是当两个系统共同存在于一个宿主中时。两个系统在 bla-IncF 质粒中存在多个匹配的识别序列（97%），表明它们是两个系统的良好靶标。此外，接合试验的结果表明，肺炎克雷伯菌中的 CRISPR-Cas 和 R-M 系统可以有效地阻止 bla 质粒的入侵。值得注意的是，CRISPR-Cas 和 R-M 系统共同作用，使 bla 质粒在接合事件中的获得减少了 4 个对数，表现出强大的协同抗质粒免疫作用。