CRISPR-Cas system positively regulates virulence of Salmonella enterica serovar Typhimurium.

IF 4.3 3区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY

Gut Pathogens Pub Date : 2024-10-26 DOI:10.1186/s13099-024-00653-5

Nandita Sharma, Ankita Das, Abhilash Vijay Nair, Palash Sethi, Vidya Devi Negi, Dipshikha Chakravortty, Sandhya Amol Marathe

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Abstract

Background: Salmonella, a foodborne pathogen, possesses a type I-E clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated (Cas) system. We investigated the system's role in regulating Salmonella virulence by deleting the CRISPR arrays and Cas operon.

Results: Our study demonstrates invasion and proliferation defects of CRISPR-Cas knockout strains in intestinal epithelial cells and macrophages owing to the repression of invasion and virulence genes. However, proliferation defects were not observed in the Gp91^phox-/- macrophages, suggesting the system's role in the pathogens' antioxidant defense. We deduced that the CRISPR-Cas system positively regulates H₂O₂ importer (OmpW), catalase (katG), peroxidase (ahpC), and superoxide dismutase (soda and sodCI), thereby protecting the cells from oxidative radicals. The knockout strains were attenuated in in-vivo infection models (Caenorhabditis elegans and BALB/c mice) due to hypersensitivity against antimicrobial peptides, complement proteins, and oxidative stress. The attenuation in virulence was attributed to the suppression of LPS modifying (pmr) genes, antioxidant genes, master regulators, and effectors of the SPI-1 (invasion) and SPI-2 (proliferation) islands in knockout strains. The regulation could be attributed to the partial complementarity of the CRISPR spacers with these genes.

Conclusions: Overall, our study extends our understanding of the role of the CRISPR-Cas system in Salmonella pathogenesis and its virulence determinants.

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CRISPR-Cas系统正向调节鼠伤寒沙门氏菌的毒力。

背景：沙门氏菌是一种食源性病原体，具有I-E型簇状规则间隔短回文重复序列（CRISPR）-CRISPR相关（Cas）系统。我们通过删除CRISPR阵列和Cas操作子研究了该系统在调节沙门氏菌毒力方面的作用：结果：我们的研究表明，CRISPR-Cas基因敲除菌株在肠上皮细胞和巨噬细胞中存在侵袭和增殖缺陷，原因是侵袭和毒力基因受到抑制。然而，在Gp91phox-/-巨噬细胞中没有观察到增殖缺陷，这表明该系统在病原体的抗氧化防御中发挥作用。我们推断 CRISPR-Cas 系统能正向调节 H2O2 输入器（OmpW）、过氧化氢酶（katG）、过氧化物酶（ahpC）和超氧化物歧化酶（soda 和 sodCI），从而保护细胞免受氧化自由基的侵害。在体内感染模型（秀丽隐杆线虫和 BALB/c 小鼠）中，基因敲除菌株由于对抗菌肽、补体蛋白和氧化应激的超敏反应而减弱。毒力减弱的原因是基因敲除菌株中的 LPS 修饰（pmr）基因、抗氧化基因、主调节因子以及 SPI-1（入侵）和 SPI-2（增殖）岛的效应因子受到抑制。这种调控可归因于CRISPR间隔与这些基因的部分互补性：总之，我们的研究拓展了我们对CRISPR-Cas系统在沙门氏菌致病过程中的作用及其毒力决定因素的认识。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Gut Pathogens GASTROENTEROLOGY & HEPATOLOGY-MICROBIOLOGY

CiteScore

7.70

自引率

2.40%

发文量

期刊介绍： Gut Pathogens is a fast publishing, inclusive and prominent international journal which recognizes the need for a publishing platform uniquely tailored to reflect the full breadth of research in the biology and medicine of pathogens, commensals and functional microbiota of the gut. The journal publishes basic, clinical and cutting-edge research on all aspects of the above mentioned organisms including probiotic bacteria and yeasts and their products. The scope also covers the related ecology, molecular genetics, physiology and epidemiology of these microbes. The journal actively invites timely reports on the novel aspects of genomics, metagenomics, microbiota profiling and systems biology. Gut Pathogens will also consider, at the discretion of the editors, descriptive studies identifying a new genome sequence of a gut microbe or a series of related microbes (such as those obtained from new hosts, niches, settings, outbreaks and epidemics) and those obtained from single or multiple hosts at one or different time points (chronological evolution).