Haider Dorina, Bauer Richard, Grempels Aline, Roscher Rebecca, Aslan Cagil Ceren, Mauerer Stefanie, Spellerberg Barbara
Institute of Medical Microbiology and Hygiene, University Hospital Ulm, Ulm, Germany.
Virulence. 2025 Dec;16(1):2541701. doi: 10.1080/21505594.2025.2541701. Epub 2025 Aug 4.
() is a commensal that can cause severe invasive bacterial infections. A considerable percentage of strains harbour CRISPR-Cas systems, which apart from being a bacterial immunity system, can play an important role in adaptation to environmental stress. The functionality of CRISPR-Cas systems has not been investigated previously. To address this, we created a set of deletion mutants in the CRISPR-Cas type II-A system of the SK52 type strain, targeting the nuclease Cas9 and CRISPR array. We were able to confirm CRISPR-Cas activity by testing these strains in a plasmid clearance assay. Furthermore, the impact of the CRISPR-Cas system on the viability of was investigated under various stress conditions, such as UV light, hydrogen peroxide exposure, and high temperatures in wild-type and CRISPR-Cas mutant strains. Under these conditions, survival was significantly lower in the strains carrying . Bacterial growth and metabolic activity in Alamar Blue assays were also negatively affected by the presence of in . In summary, we found that the presence of a functional CRISPR-Cas system in leads to measurable metabolic and fitness costs in the wild-type strain. Carrying was associated with an impaired stress response in our experiments and may explain why many strains of this species lack CRISPR-Cas.
(某细菌)是一种共生菌,可引起严重的侵袭性细菌感染。相当比例的该细菌菌株含有CRISPR-Cas系统,该系统除作为细菌免疫系统外,在适应环境压力方面也可发挥重要作用。此前尚未对该细菌的CRISPR-Cas系统功能进行研究。为解决这一问题,我们在SK52型菌株的CRISPR-Cas II-A型系统中创建了一组缺失突变体,靶向核酸酶Cas9和CRISPR阵列。我们通过在质粒清除试验中测试这些菌株,证实了CRISPR-Cas活性。此外,还在野生型该细菌和CRISPR-Cas突变株中,研究了CRISPR-Cas系统在紫外线、过氧化氢暴露和高温等各种应激条件下对该细菌生存能力的影响。在这些条件下,携带(相关基因或元件)的菌株存活率显著降低。Alamar Blue试验中的细菌生长和代谢活性也受到该细菌中(相关基因或元件)存在的负面影响。总之,我们发现该细菌中功能性CRISPR-Cas系统的存在会导致野生型菌株出现可测量的代谢和适应性代价。在我们的实验中,携带(相关基因或元件)与应激反应受损有关,这可能解释了为什么该物种的许多菌株缺乏CRISPR-Cas。