Di Cianni Nadia, Bolsinger Simon, Brendel Jutta, Raabe Monika, König Sabine, Mitchell Laura, Bischler Thorsten, Gräfenhan Tom, Read Clarissa, Erdmann Susanne, Allers Thorsten, Walther Paul, Urlaub Henning, Dyall-Smith Mike, Pfeiffer Friedhelm, Marchfelder Anita
Molecular Biology and Biotechnology of Prokaryotes, Ulm University, Ulm 89069, Germany.
Max Planck Institute for Multidisciplinary Sciences, Bioanalytical Mass Spectrometry Group, Göttingen 37077, Germany.
Microlife. 2025 May 19;6:uqaf008. doi: 10.1093/femsml/uqaf008. eCollection 2025.
harbours four putative proviruses: Halfvol1, Halfvol2, Halfvol3, and Halfvol4. In this study, we successfully deleted all four provirus genomes, demonstrating, that they are not essential. Transcriptome comparison between this strain (∆Halfvol1-4) and a wild-type strain reveals an increase in archaella and chemotaxis gene expression, resulting in higher swarming motility in ∆Halfvol1-4. Furthermore, ∆Halfvol1-4 cells show an elongated cell shape and a higher resistance to HO stress compared to the wild type. RNA-seq also revealed downregulation of CRISPR arrays in the provirus-free strain. Circularised genomes of Halfvol1, Halfvol2, and Halfvol3 were found in the culture supernatant of the wild-type strain. This confirms excision of the proviruses from the chromosome, which seems to happen more efficiently at low temperature (30°C). Electron microscopy revealed potential viral particles in the supernatant, and mass spectrometry analysis confirmed the presence of structural viral proteins of Halfvol1 and Halfvol3 in the isolated virus sample. These observations suggest that these proviruses are active and cause a chronic infection in .
Halfvol1、Halfvol2、Halfvol3和Halfvol4。在本研究中,我们成功删除了所有四种前病毒基因组,证明它们并非必需。该菌株(∆Halfvol1-4)与野生型菌株之间的转录组比较显示,古菌鞭毛和趋化性基因表达增加,导致∆Halfvol1-4的群体运动性更高。此外,与野生型相比,∆Halfvol1-4细胞呈现出细长的细胞形状,对HO应激具有更高的抗性。RNA测序还显示无前病毒菌株中CRISPR阵列的下调。在野生型菌株的培养上清液中发现了Halfvol1、Halfvol2和Halfvol3的环状基因组。这证实了前病毒从染色体上的切除,这似乎在低温(30°C)下更有效地发生。电子显微镜显示上清液中存在潜在的病毒颗粒,质谱分析证实分离的病毒样品中存在Halfvol1和Halfvol3的结构病毒蛋白。这些观察结果表明,这些前病毒具有活性,并在……中引起慢性感染。