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(A)/(D) 携带的链球菌噬菌体 Φ1207.3 编码了一个 SOS 样系统,该系统由 UV-C 光诱导,负责提高生存能力和增加突变率。

The (A)/(D)-carrying streptococcal prophage Φ1207.3 encodes an SOS-like system, induced by UV-C light, responsible for increased survival and increased mutation rate.

机构信息

Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena , Siena, Italy.

Unidad Clínica de Enfermedades Infecciosas, Microbiología y Medicina Preventiva, Hospital Universitario Virgen Macarena , Sevilla, Spain.

出版信息

J Bacteriol. 2023 Sep 26;205(9):e0019123. doi: 10.1128/jb.00191-23. Epub 2023 Sep 12.

Abstract

Bacterial SOS response is an inducible system of DNA repair and mutagenesis. Streptococci lack a canonical SOS response, but an SOS-like response was reported in some species. The (A)-(D)-carrying prophage Ф1207.3 of contains a region, spanning to , showing homology to characterized streptococcal SOS-like cassettes. Genome-wide homology search showed the presence of the whole Φ1207.3 SOS-like cassette in three . prophages, while parts of it were found in other bacterial species. To investigate whether this cassette confers an SOS-mutagenesis phenotype, we constructed R6 isogenic derivative strains: (i) FR172, streptomycin resistant, (ii) FR173, carrying Φ1207.3, and (iii) FR174, carrying a recombinant Φ1207.3, where the SOS-like cassette was deleted. These strains were used in survival and mutation rate assays using a UV-C LED instrument, for which we designed and 3D-printed a customized equipment, constituted of an instrument support and swappable-autoclavable mini-plates and lids. Upon exposure to UV fluences ranging from 0 to 6,400 J/m at four different wavelengths, 255, 265, 275, and 285 nm, we found that the presence of Φ1207.3 SOS-like cassette increases bacterial survival up to 34-fold. Mutation rate was determined by measuring rifampicin resistance acquisition upon exposure to UV fluence of 50 J/m at the four wavelengths by fluctuation test. The presence of Φ1207.3 SOS-like cassette resulted in a significant increase in the mutation rate (up to 18-fold) at every wavelength. In conclusion, we demonstrated that Φ1207.3 carries a functional SOS-like cassette responsible for an increased survival and increased mutation rate in . IMPORTANCE Bacterial mutation rate is generally low, but stress conditions and DNA damage can induce stress response systems, which allow for improved survival and continuous replication. The SOS response is a DNA repair mechanism activated by some bacteria in response to stressful conditions, which leads to a temporary hypermutable phenotype and is usually absent in streptococcal genomes. Here, using a reproducible and controlled UV irradiation system, we demonstrated that the SOS-like gene cassette of prophage Φ1207.3 is functional, responsible for a temporary hypermutable phenotype, and enhances bacterial survival to UV irradiation. Prophage Φ1207.3 also carries erythromycin resistance genes and can lysogenize different pathogenic bacteria, constituting an example of a mobile genetic element which can confer multiple phenotypes to its host.

摘要

细菌 SOS 反应是一种诱导型的 DNA 修复和诱变系统。链球菌缺乏典型的 SOS 反应,但在一些物种中报道了 SOS 样反应。携带 (A)-(D) 的噬菌体 Ф1207.3 包含一个区域,跨度为 到 ,与已鉴定的链球菌 SOS 样盒式有关。全基因组同源性搜索显示,三个 中存在整个 Ф1207.3 SOS 样盒式,而其他细菌物种中则存在其部分。为了研究这个盒式是否赋予了 SOS 诱变表型,我们构建了 6 个 R 同源衍生菌株:(i) FR172,链霉素抗性,(ii) FR173,携带 Ф1207.3,和 (iii) FR174,携带重组 Ф1207.3,其中 SOS 样盒式缺失。我们使用紫外线-C LED 仪器进行了生存和突变率测定实验,为此我们设计并 3D 打印了一个定制设备,由仪器支架和可更换的高压灭菌迷你板和盖子组成。在四个不同波长 255、265、275 和 285nm 下,我们发现紫外线剂量从 0 到 6400J/m 时,存在 Ф1207.3 SOS 样盒式会使细菌的存活率提高 34 倍。通过在四个波长下的紫外线剂量为 50J/m 时进行波动试验,测定 rifampicin 抗性获得率来确定突变率。Ф1207.3 SOS 样盒式的存在导致突变率显著增加(高达 18 倍),在每个波长下均如此。总之,我们证明 Ф1207.3 携带一个功能性的 SOS 样盒式,负责提高 的生存能力和提高突变率。 重要性 细菌的突变率通常较低,但压力条件和 DNA 损伤可以诱导应激反应系统,从而提高生存能力和持续复制能力。SOS 反应是一种由一些细菌在应激条件下激活的 DNA 修复机制,导致暂时的超突变表型,通常不存在于链球菌基因组中。在这里,我们使用可重复和可控的紫外线照射系统,证明了噬菌体 Ф1207.3 的 SOS 样基因盒式是有功能的,负责暂时的超突变表型,并增强了细菌对紫外线照射的生存能力。噬菌体 Ф1207.3 还携带红霉素抗性基因,可溶原化不同的致病菌,构成了一个可向宿主赋予多种表型的移动遗传元件的例子。

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