To test the effect of humidity on the radiation resistance of Deinococcus radiodurans, air-dried cells were irradiated with germicidal 254 nm UV, and simulated environmental UV or γ-radiation and survival was compared to cells in suspension. It was observed that desiccated cells exhibited higher levels of resistance than cells in suspension toward UV or γ-radiation as well as after 85°C heat shock. It was also shown that low relative humidity improves survival during long-term storage of desiccated D. radiodurans cells. It can be concluded that periods or environments in which cells exist in a dehydrated state are beneficial for D. radiodurans' survival exposed to various other stresses.
To study the role of different DNA repair genes in the resistance of Deinococcus radiodurans to mono- and polychromatic UV radiation, wild-type strain and knockout mutants in RecA, PprA, and IrrE of D. radiodurans were irradiated with UV-C (254 nm), UV-(A + B) (280-400 nm) and UV-A (315-400 nm) radiation, and survival was monitored. The strain deficient in recA was highly sensitive to UV-C radiation compared to the wild-type, but showed no loss of resistance against irradiation with UV-(A + B) and UV-A, while pprA and irrE-deficient strains exhibited elevated sensitivity to UV-A and UV-(A + B) radiation. These results suggest that the repair of DNA double-strand breaks is essential after treatment with highly energetic UV-C radiation, whereas protection from oxidative stress may play a greater role in resistance to environmentally relevant UV radiation.
To study the effects of heat shock on Deinococcus radiodurans and the role of DNA repair in high temperature resistance, different strains of D. radiodurans (wild type, recA, irrE, and pprA) were treated with temperatures ranging from 40 to 100 °C under wet and dry conditions. The mutant strains were more sensitive to wet heat of ≥60 °C and dry heat of ≥80 °C than the wild type. Both wild-type and DNA repair-deficient strains were much more resistant to high temperatures when exposed in the dried state as opposed to cells in suspension. Molecular staining techniques with the wild-type strain revealed that cells in the dried state were able to retain membrane integrity after drying and subsequent heat exposure, while heat-exposed cells in suspension showed significant loss of membrane integrity and respiration activity. The results suggest that the repair of DNA damage (e.g., DNA double-strand breaks by RecA and PprA) is essential after treatment with wet heat at temperatures >60 °C and dry heat >80 °C, and the ability of D. radiodurans to stabilize its plasma membrane during dehydration might represent one aspect in the protection of dried cells from heat-induced membrane damage.
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