Department of Clinical Medicine and Surgery, University of Naples Federico II, 81031 Naples, Italy.
Department of Plant Pathology, Division for Phytomedicine, Faculty of Agriculture, University of Zagreb, 10000 Zagreb, Croatia.
Int J Mol Sci. 2022 Sep 18;23(18):10917. doi: 10.3390/ijms231810917.
Bacterial SSB proteins, as well as their eukaryotic RPA analogues, are essential and ubiquitous. They avidly bind single-stranded DNA and regulate/coordinate its metabolism, hence enabling essential DNA processes such as replication, transcription, and repair. The prototypic SSB protein is encoded by an gene. Although the gene promoters harbor an SOS box, multiple studies over several decades failed to elucidate whether gene expression is inducible and SOS dependent. The SOS regulon is comprised of about 50 genes, whose transcription is coordinately induced under stress conditions. Using quantitative real-time PCR, we determined the gene expression kinetics in UV- and γ-irradiated and revealed that gene expression is elevated in irradiated cells in an SOS-dependent manner. Additionally, the expression of the gene was determined to indicate the extent of SOS induction. In a mutant with a constitutively induced SOS regulon, the gene was overexpressed in the absence of DNA damage. Furthermore, we measured gene expression by droplet digital PCR during unaffected bacterial growth and revealed that gene expression was equal in wild-type and SOS bacteria, whereas expression was higher in the former. This study thus reveals a complex pattern of gene expression, which under stress conditions depends on the SOS regulon, whereas during normal bacterial growth it is unlinked to SOS induction. The gene is SOS regulated in such a way that its basal expression is relatively high and can be increased only through stronger SOS induction. The remarkable SOS induction observed in undisturbed wild-type cells may challenge our notion of the physiological role of the SOS response in bacteria.
细菌 SSB 蛋白及其真核 RPA 类似物是必不可少且普遍存在的。它们能够强烈地结合单链 DNA,并调节/协调其代谢,从而使复制、转录和修复等基本 DNA 过程成为可能。典型的 SSB 蛋白由 基因编码。尽管 基因启动子含有 SOS 盒,但几十年来的多项研究未能阐明 基因表达是否可诱导且依赖 SOS。SOS 调控子由大约 50 个基因组成,其转录在应激条件下协调诱导。我们使用定量实时 PCR 测定了 UV 和 γ 辐射 中的 基因表达动力学,并发现 基因表达在 SOS 依赖的方式下在辐照细胞中升高。此外, 基因的表达被确定为指示 SOS 诱导程度的指标。在一个具有组成型诱导的 SOS 调控子的突变体中,即使没有 DNA 损伤, 基因也会过表达。此外,我们通过液滴数字 PCR 在不受影响的细菌生长期间测量了 基因表达,并发现 基因在野生型和 SOS 细菌中的表达相等,而在前者中的表达更高。因此,这项研究揭示了 基因表达的复杂模式,在应激条件下,其表达依赖于 SOS 调控子,而在正常细菌生长过程中,其表达与 SOS 诱导无关。 基因以这样的方式受到 SOS 调控,即其基础表达相对较高,并且只能通过更强的 SOS 诱导来增加。在未受干扰的野生型细胞中观察到的显著 SOS 诱导可能会挑战我们对细菌中 SOS 反应生理作用的认识。