Institute of Bio- and Geosciences: IBG-1, Forschungszentrum Jülich, 52425 Jülich, Germany.
Viruses. 2021 Mar 17;13(3):495. doi: 10.3390/v13030495.
In this study, we provide a comprehensive analysis of the genomic features of the phage CL31 and the infection dynamics with the biotechnologically relevant host strain ATCC 13032. Genome sequencing and annotation of CL31 revealed a 45-kbp genome composed of 72 open reading frames, mimicking the GC content of its host strain (54.4%). An ANI-based distance matrix showed the highest similarity of CL31 to the temperate corynephage Φ16. While the ATCC 13032 wild type strain showed only mild propagation of CL31, a strain lacking the -- restriction-modification system was efficiently infected by this phage. Interestingly, the prophage-free strain MB001 featured an even accelerated amplification of CL31 compared to the ∆ strain suggesting a role of cryptic prophage elements in phage defense. Proteome analysis of purified phage particles and transcriptome analysis provide important insights into structural components of the phage and the response of to CL31 infection. Isolation and sequencing of CL31-resistant strains revealed SNPs in genes involved in mycolic acid biosynthesis suggesting a role of this cell envelope component in phage adsorption. Altogether, these results provide an important basis for further investigation of phage-host interactions in this important biotechnological model organism.
在这项研究中,我们对噬菌体 CL31 的基因组特征及其与生物技术相关宿主菌株 ATCC 13032 的感染动力学进行了全面分析。CL31 的基因组测序和注释揭示了一个由 72 个开放阅读框组成的 45-kbp 基因组,其 GC 含量与宿主菌株相似(54.4%)。基于 ANI 的距离矩阵显示,CL31 与温和的棒状噬菌体 Φ16 最为相似。尽管 ATCC 13032 野生型菌株对 CL31 的繁殖仅表现出轻微的影响,但缺乏 -- 限制修饰系统的菌株却能被该噬菌体有效感染。有趣的是,与 ∆ 菌株相比,不含原噬菌体的 MB001 菌株甚至能加速 CL31 的扩增,这表明潜伏原噬菌体元件在噬菌体防御中发挥作用。纯化噬菌体颗粒的蛋白质组分析和转录组分析为噬菌体的结构成分和宿主对 CL31 感染的反应提供了重要的见解。CL31 抗性菌株的分离和测序揭示了参与分枝菌酸生物合成的基因中的 SNPs,表明该细胞包膜成分在噬菌体吸附中发挥作用。总之,这些结果为进一步研究这个重要的生物技术模式生物中的噬菌体-宿主相互作用提供了重要的基础。