FG11 Division of Enteropathogenic Bacteria and Legionella, Robert Koch Institute, 38855 Wernigerode, Germany.
MF1 Bioinformatic Support, Robert Koch Institute, 13353 Berlin, Germany.
Genome Res. 2022 Sep 27;32(9):1711-1726. doi: 10.1101/gr.276747.122.
The Gram-positive bacterium occurs widespread in the environment and infects humans when ingested along with contaminated food. Such infections are particularly dangerous for risk group patients, for whom they represent a life-threatening disease. To invent novel strategies to control contamination and disease, it is important to identify those cellular processes that maintain pathogen growth inside and outside the host. Here, we have applied transposon insertion sequencing (Tn-Seq) to for the identification of such processes on a genome-wide scale. Our approach identified 394 open reading frames that are required for growth under standard laboratory conditions and 42 further genes, which become necessary during intracellular growth in macrophages. Most of these genes encode components of the translation machinery and act in chromosome-related processes, cell division, and biosynthesis of the cellular envelope. Several cofactor biosynthesis pathways and 29 genes with unknown functions are also required for growth, suggesting novel options for the development of antilisterial drugs. Among the genes specifically required during intracellular growth are known virulence factors, genes compensating intracellular auxotrophies, and several cell division genes. Our experiments also highlight the importance of PASTA kinase signaling for general viability and of glycine metabolism and chromosome segregation for efficient intracellular growth of .
革兰氏阳性菌 广泛存在于环境中，当与污染食物一起摄入时会感染人类。对于高危人群患者来说，此类感染尤其危险，因为它们可能导致危及生命的疾病。为了发明新的策略来控制污染和疾病，识别那些在宿主内外维持病原体生长的细胞过程非常重要。在这里，我们应用转座子插入测序 (Tn-Seq) 技术在全基因组范围内鉴定这些过程。我们的方法鉴定了 394 个开放阅读框，这些基因是在标准实验室条件下生长所必需的，还有 42 个进一步的基因，这些基因在巨噬细胞内的生长过程中是必需的。这些基因中的大多数编码翻译机制的组成部分，并参与染色体相关过程、细胞分裂和细胞包膜的生物合成。几种辅助因子生物合成途径和 29 个具有未知功能的基因也需要生长，这为开发抗李斯特菌药物提供了新的选择。在细胞内生长过程中特别需要的基因包括已知的毒力因子、补偿细胞内营养缺陷的基因和几个细胞分裂基因。我们的实验还强调了 PASTA 激酶信号对一般生存能力的重要性，以及甘氨酸代谢和染色体分离对 有效细胞内生长的重要性。
