Ye Qianying, Shin Eunjung, Lee Chanjae, Choi Nakjun, Kim Yeonho, Yoon Ki Sun, Lee Sung-Jae
Department of Biology and Nutrition, Kyung Hee University, Seoul 02447, Republic of Korea.
Department of Food and Nutrition, Kyung Hee University, Seoul 02447, Republic of Korea.
J Microbiol Methods. 2022 May;196:106473. doi: 10.1016/j.mimet.2022.106473. Epub 2022 Apr 22.
Insertion sequences (ISs) of the radiation-resistant bacterium Deinococcus geothermalis are transposed into other loci by oxidative stress through hydrogen peroxide treatment. Gamma irradiation and dielectric barrier discharge (DBD) plasma radiation are known to produce a variety of oxidative stress agents such as reactive oxygen species and reactive nitrogen species. Therefore, to determine whether the transposition of ISs was induced in D. geothermalis by both gamma irradiation and DBD plasma radiation, we selected non-pigmented mutants with disrupted target genes encoding carotenoid biosynthesis enzymes such as a phytoene synthase (dgeo_0523) and a phytoene desaturase (dgeo_0524). Different DNA-binding protein-deficient mutants exhibited novel transposition of ISs. Dps (dgeo_0257), OxyR (dgeo_1888), and the LysR (dgeo_2840) family regulator, in addition to cystine importer-disrupted and -overexpressed mutants (dgeo_1986-87 and dgeo_1985R) and wild-type D. geothermalis were tested in this study. Active IS transposition was not detected in two wild-type control species (Deinococcus radiodurans and Deinococcus radiopugnans) after phenotypic selection in gamma irradiation. Our finding demonstrated that gamma irradiation triggers the transposition of particular IS elements, especially ISDge2 and ISDge3 of the IS1 family, ISDge5 of the IS701 family, and ISDge6 of the IS5 family in wild-type strain and the Δdgeo_0257, Δdgeo_1986-87, Δdgeo_1985R, and Δdgeo_2840 mutants. Furthermore, DBD plasma radiation triggered the transposition of ISDge11 of the IS4 family in the wild-type strain; ISDge6 of the IS5 family on Δdgeo_0257, Δdgeo_1888 and Δdgeo_2840; ISDge5 of the IS701 family on Δdgeo_0257 strain.
抗辐射细菌嗜热栖热放线菌的插入序列(ISs)通过过氧化氢处理产生的氧化应激被转座到其他基因座。已知γ射线辐照和介质阻挡放电(DBD)等离子体辐射会产生多种氧化应激因子,如活性氧和活性氮。因此,为了确定γ射线辐照和DBD等离子体辐射是否会在嗜热栖热放线菌中诱导ISs的转座,我们选择了编码类胡萝卜素生物合成酶(如八氢番茄红素合酶(dgeo_0523)和八氢番茄红素去饱和酶(dgeo_0524))的靶基因被破坏的无色素突变体。不同的DNA结合蛋白缺陷突变体表现出ISs的新转座。本研究测试了Dps(dgeo_0257)、OxyR(dgeo_1888)和LysR(dgeo_2840)家族调节因子,以及胱氨酸导入缺陷和过表达突变体(dgeo_1986 - 87和dgeo_1985R)和野生型嗜热栖热放线菌。在γ射线辐照下进行表型选择后,在两个野生型对照菌株(耐辐射球菌和放射抗性球菌)中未检测到活跃的IS转座。我们的研究结果表明,γ射线辐照会触发野生型菌株以及Δdgeo_0257、Δdgeo_1986 - 87、Δdgeo_1985R和Δdgeo_2840突变体中特定IS元件的转座,特别是IS1家族的ISDge2和ISDge3、IS701家族的ISDge5以及IS5家族的ISDge6。此外,DBD等离子体辐射会触发野生型菌株中IS4家族的ISDge11的转座;在Δdgeo_0257、Δdgeo_1888和Δdgeo_2840中IS5家族的ISDge6的转座;在Δdgeo_0257菌株中IS701家族的ISDge5的转座。