Department of Science and Engineering for Sustainable Development, Faculty of Science and Engineering, Soka University.
Microbes Environ. 2019 Dec 27;34(4):363-373. doi: 10.1264/jsme2.ME19055. Epub 2019 Sep 21.
Several species of Sulfolobales have been used as model organisms in the study of response mechanisms to ultraviolet (UV) irradiation in hyperthermophilic crenarchaea. To date, the transcriptional responses of genes involved in the initiation of DNA replication, transcriptional regulation, protein phosphorylation, and hypothetical function have been observed in Sulfolobales species after UV irradiation. However, due to the absence of knockout experiments, the functions of these genes under in situ UV irradiation have not yet been demonstrated. In the present study, we constructed five gene knockout strains (cdc6-2, tfb3, rio1, and two genes encoding the hypothetical proteins, Saci_0951 and Saci_1302) of Sulfolobus acidocaldarius and examined their sensitivities to UV irradiation. The knockout strains exhibited significant sensitivities to UV-B irradiation, indicating that the five UV-regulated genes play an important role in responses to UV irradiation in vivo. Furthermore, Δcdc6-2, Δrio1, ΔSaci_0951, and Δtfb3 were sensitive to a wide variety of helix-distorting DNA lesions, including UV-induced DNA damage, an intra-strand crosslink, and bulky adducts. These results reveal that cdc6-2, tfb3, rio1, and Saci_0951 are play more important roles in broad responses to helix-distorting DNA damage than in specific responses to UV irradiation.
几种 Sulfolobales 物种已被用作研究嗜热古菌中紫外线 (UV) 照射响应机制的模式生物。迄今为止,在 Sulfolobales 物种中观察到了与 DNA 复制起始、转录调控、蛋白磷酸化和假设功能相关的基因的转录响应。然而,由于缺乏敲除实验,这些基因在原位 UV 照射下的功能尚未得到证明。在本研究中,我们构建了 Sulfolobus acidocaldarius 的五个基因敲除菌株(cdc6-2、tfb3、rio1 和两个编码假设蛋白 Saci_0951 和 Saci_1302 的基因),并研究了它们对 UV 照射的敏感性。敲除菌株对 UV-B 照射表现出显著的敏感性,表明这五个 UV 调节基因在体内对 UV 照射的响应中发挥着重要作用。此外,Δcdc6-2、Δrio1、ΔSaci_0951 和 Δtfb3 对多种螺旋扭曲 DNA 损伤敏感,包括 UV 诱导的 DNA 损伤、链内交联和大体积加合物。这些结果表明,cdc6-2、tfb3、rio1 和 Saci_0951 在广泛响应螺旋扭曲 DNA 损伤方面比在特定响应 UV 照射方面发挥更重要的作用。