Delgado-Nungaray Javier Alejandro, Figueroa-Yáñez Luis Joel, Reynaga-Delgado Eire, Corona-España Ana Montserrat, Gonzalez-Reynoso Orfil
Chemical Engineering Department, University Center for Exact and Engineering Sciences, University of Guadalajara, Guadalajara, Jalisco, Mexico.
Industrial Biotechnology Unit, Center for Research and Assistance in Technology and Design of the State of Jalisco, A.C. (CIATEJ), Guadalajara, Jalisco, Mexico.
PLoS One. 2024 Dec 31;19(12):e0312783. doi: 10.1371/journal.pone.0312783. eCollection 2024.
Multidrug resistance in Pseudomonas aeruginosa, a high-priority pathogen per the World Health Organization, poses a global threat due to carbapenem resistance and limited antibiotic treatments. Using the bioinformatic tools CRISPRCasFinder, CRISPRCasTyper, CRISPRloci, and CRISPRImmunity, we analyzed the genome of P. aeruginosa PAO1 and revealed an orphan CRISPR system, suggesting it may be a remnant of a type IV system due to the presence of the DinG protein. This system comprises two CRISPR arrays and noteworthy DinG and Cas3 proteins, supporting recent evidence about the association between type IV and I CRISPR systems. Additionally, we demonstrated a co-evolutionary relationship between the orphan CRISPR system in P. aeruginosa PAO1 and the mobile genetic element and prophages identified. One self-targeting spacer was identified, often associated with bacterial evolution and autoimmunity, and no Acr proteins. This research opens avenues for studying how these CRISPR arrays regulate pathogenicity and for developing alternative strategies using its endogenous orphan CRISPR system against carbapenem-resistant P. aeruginosa strains.
铜绿假单胞菌是世界卫生组织认定的重点病原体，其多重耐药性因对碳青霉烯类药物耐药且抗生素治疗手段有限而构成全球威胁。我们使用生物信息学工具CRISPRCasFinder、CRISPRCasTyper、CRISPRloci和CRISPRImmunity对铜绿假单胞菌PAO1的基因组进行了分析，发现了一个孤儿CRISPR系统，由于存在DinG蛋白，表明它可能是IV型系统的残余。该系统由两个CRISPR阵列以及值得注意的DinG和Cas3蛋白组成，支持了关于IV型和I型CRISPR系统之间关联的最新证据。此外，我们证明了铜绿假单胞菌PAO1中的孤儿CRISPR系统与所鉴定的移动遗传元件和原噬菌体之间存在共同进化关系。鉴定出了一个自我靶向间隔序列，其通常与细菌进化和自身免疫相关，且未发现Acr蛋白。这项研究为研究这些CRISPR阵列如何调节致病性以及利用其内源性孤儿CRISPR系统针对耐碳青霉烯类铜绿假单胞菌菌株开发替代策略开辟了道路。
