Kretz Jonas, Börner Janek, Friedrich Tobias, McIntosh Matthew, Procida-Kowalski Tara, Gerken Florian, Wilhelm Jochen, Klug Gabriele
Institute of Microbiology and Molecular Biology, Justus-Liebig-University, Giessen, Germany.
Institute of Biochemistry, Justus-Liebig-University, Giessen, Germany.
Front Microbiol. 2024 Apr 29;15:1384543. doi: 10.3389/fmicb.2024.1384543. eCollection 2024.
Bacteria use CRISPR Cas systems to defend against invading foreign nucleic acids, e.g., phage genomes, plasmids or mobile genetic elements. Some CRISPR Cas systems were reported to have physiological importance under a variety of abiotic stress conditions. We used physiological tests under different stress conditions and RNA-seq analyses to address the possible function of the RNA-targeting class 2 type VI CRISPR Cas system of the facultative phototrophic α-proteobacterium . Expression of the system was low under exponential non-stress conditions and high during oxidative stress, membrane stress and in stationary phase. Induction of the CRISPR Cas system in presence of a target protospacer RNA resulted in a growth arrest of . RNA-seq revealed a strong alteration of the transcriptome when was induced in presence of a target protospacer. RNA 5' end mapping indicated that the CRISPR Cas-dependent transcriptome remodeling is accompanied by fragmentation of cellular RNAs, e.g., for mRNAs originating from a genomic locus which encodes multiple ribosomal proteins and the RNA polymerase subunits RpoA, RpoB and RpoC. The data suggest a function of this CRISPR Cas system in regulated growth arrest, which may prevent the spread of phages within the population.
细菌利用CRISPR Cas系统抵御入侵的外源核酸,例如噬菌体基因组、质粒或移动遗传元件。据报道,一些CRISPR Cas系统在多种非生物胁迫条件下具有生理重要性。我们通过在不同胁迫条件下的生理测试和RNA测序分析,来探究兼性光合α-变形杆菌中靶向RNA的2类VI型CRISPR Cas系统的可能功能。该系统在指数生长的非胁迫条件下表达水平较低,而在氧化应激、膜应激和稳定期时表达水平较高。在存在靶标原间隔序列RNA的情况下诱导CRISPR Cas系统会导致[具体细菌名称未给出]生长停滞。RNA测序显示,当在存在靶标原间隔序列的情况下诱导[具体细菌名称未给出]时,转录组发生了强烈改变。RNA 5'末端图谱分析表明,依赖CRISPR Cas的转录组重塑伴随着细胞RNA的片段化,例如,对于源自一个编码多种核糖体蛋白以及RNA聚合酶亚基RpoA、RpoB和RpoC的基因组位点的mRNA。这些数据表明该CRISPR Cas系统在调控生长停滞中发挥作用,这可能会阻止噬菌体在群体中的传播。