Abstract
The transposition of insertion sequence elements was evaluated among different Deinococcus geothermalis lineages, including the wild-type, a cystine importer-disrupted mutant, a complemented strain, and a cystine importer-overexpressed strain. Cellular growth reached early exponential growth at OD600 2.0 and late exponential growth at OD600 4.0. Exposing the cells to hydrogen peroxide (80–100 mM) resulted in the transposition of insertion sequences (ISs) in genes associated with the carotenoid biosynthesis pathway. Particularly, ISDge7 (an IS5 family member) and ISDge5 (an IS701 family member) from the cystine importer-disrupted mutant were transposed into phytoene desaturase (dgeo_0524) via replicative transposition. Further, the cystine importer-overexpressed strain Δdgeo_1985R showed transposition of both ISDge2 and ISDge5 elements. In contrast, IS transposition was not detected in the complementary strain. Interestingly, a cystine importer-overexpressing strain exhibited streptomycin resistance, indicating that point mutation occurred in the rpsL (dgeo_1873) gene encoding ribosomal protein S12. qRT-PCR analyses were then conducted to evaluate the expression of oxidative stress response genes, IS elements, and low-molecular-weight thiol compounds such as mycothiol and bacillithiol. Nevertheless, the mechanisms that trigger IS transposition in redox imbalance conditions remain unclear. Here, we report that the active transposition of different IS elements was affected by intracellular redox imbalances caused by cystine importer deficiencies or overexpression.
Highlights
IntroductionRedox balance is largely regulated by the uptake of cysteine and cystine-derived thiol compounds from the environment through membrane channels, as well as the activity of specific importers
All experimental strains were confirmed via Polymerase chain reaction (PCR) detection (Figure 1)
For “redox biology” because it is an essential process for living organisms, which evolved from signalling processes in many physiological contexts [38]
Summary
Redox balance is largely regulated by the uptake of cysteine and cystine-derived thiol compounds from the environment through membrane channels, as well as the activity of specific importers. This mechanism maintains the intracellular redox balance in Gram-positive and Gram-negative bacteria [3,4,5]. Reactive oxygen species (ROS), including H2 O2 , are known to alter protein structure, resulting in the activation of catalytic sites via the oxidation of cysteine residues, in addition to enzyme inactivation and DNA damage [6,7]. Many types of thiol compounds are known to counteract the adverse effects of oxidative stress caused by ROS such as hydrogen peroxide, hydroxyl radicals, and superoxide
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