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The role of DNA repair by nonhomologous-end joining (NHEJ) in spore resistance to UV, ionizing radiation, and ultrahigh vacuum was studied in wild-type and DNA repair mutants (recA, splB, ykoU, ykoV, and ykoU ykoV mutants) of Bacillus subtilis. NHEJ-defective spores with mutations in ykoU, ykoV, and ykoU ykoV were significantly more sensitive to UV, ionizing radiation, and ultrahigh vacuum than wild-type spores, indicating that NHEJ provides an important pathway during spore germination for repair of DNA double-strand breaks.It has been shown that endospores of gram-positive bacteria can remain viable for at least thousands of years (5, 44, 54; reviewed in reference 31). Bacterial spores persist in a metabolically inactive state, and environmental damage to spore cellular components accumulates unrepaired until germination and outgrowth (32). However, Bacillus subtilis spores are highly resistant to different environmental stresses, such as toxic chemicals and biocidal agents, desiccation, pressure and temperature extremes, and ionizing and UV radiation. The reason for this high resistance to environmental extremes lies partly in the spore structure itself: spores possess thick layers of highly cross-linked coat proteins (13), a modified peptidoglycan spore cortex, and abundant intracellular constituents such as the calcium chelate of dipicolinic acid and small, acidsoluble spore proteins (␣/␤-type SASP), as protectants of spore DNA (46, 50). Binding of ␣/␤-type SASP to spore DNA, coupled with spore core dehydration, appears to change the helical conformation of spore DNA from the B form to an A-like form (34, 48), which in turn alters its UV photochemistry to favor the production of 5-thyminyl-5,6-dihydrothymine, the unique spore-specific spore photoproduct (SP) (8,32,35,50). For the removal of the SP, spores possess an SP-specific repair enzyme called SP lyase, encoded by the splB gene, that monomerizes the SP dimer back to two thymine residues in an adenosyl-radical-dependent reaction (4,28,42).While the UV photochemistry of spore DNA and the repair of UV damage to DNA during germination are well described (12,32,33,47,50), there has been relatively little work on the nature of DNA damage in spores caused by ionizing radiation or extreme dryness and on the occurrence of a specific DNA repair system(s) for repair of this damage. It is assumed that DNA double-strand breaks (DSB), which are the most critical damage caused by ionizing radiation (57) and desiccation (9, 10, 39) in vegetative cells, are also induced in bacterial spores. Spores of B. subtilis contain a single chromosome arranged in a toroidal shape (16, 41); therefore, the homologous recombination pathway, which requires at least two homologous chromosomes, cannot operate on DSB during spore germination (55). An alternative repair pathway for DSB induced in spore DNA, nonhomologous-end joining (NHEJ) (3, 56), is considered here. This pathway as it occurs in eukaryotic cells requires a DNA end-binding component called Ku (Ku70 and Ku80) (58). The fir...

The role of DNA repair by nonhomologous end joining (NHEJ), homologous recombination, spore photoproduct lyase, and DNA polymerase I and genome protection via ␣/␤-type small, acid-soluble spore proteins (SASP) in Bacillus subtilis spore resistance to accelerated heavy ions (high-energy charged [HZE] particles) and X rays has been studied. Spores deficient in NHEJ and ␣/␤-type SASP were significantly more sensitive to HZE particle bombardment and X-ray irradiation than were the recA, polA, and splB mutant and wild-type spores, indicating that NHEJ provides an efficient DNA double-strand break repair pathway during spore germination and that the loss of the ␣/␤-type SASP leads to a significant radiosensitivity to ionizing radiation, suggesting the essential function of these spore proteins as protectants of spore DNA against ionizing radiation.Endospores of the gram-positive bacterium Bacillus subtilis are highly resistant to inactivation by environmental stresses, such as biocidal agents and toxic chemicals, desiccation, pressure and temperature extremes, and high fluences of UV radiation (reviewed in references 44, 45, and 61) and are a powerful biodosimetric system for terrestrial environmental monitoring and astrobiological studies (44). On Earth, understanding extreme spore resistance to ionizing radiation is important in the areas of food preservation, medical sterilization, and decontamination from bioterror attack (4, 17, 47; reviewed in references 42 and 43). Off Earth, spore radiation resistance is important both in space flight and in ground-based simulations, in order to obtain information on the biological damage produced by exposure to space conditions (23,25,26,44). Onboard several spacecraft (Apollo 16, Spacelab 1, LDEF, D2, and FOTON), spores of Bacillus subtilis were exposed to selected parameters of space, such as space vacuum and different spectral ranges of solar UV radiation and cosmic rays, applied separately or in combination (5, 9, 19-21, 23, 24, 26). Especially, the radiation environment on Earth, on Mars, in low-Earth orbit, and in deep space is typified by a wide variety of primary particles covering an extended range of energies. Galactic cosmic rays (GCR) are charged particles that originate from sources beyond our solar system. The distribution of GCR is believed to be isotropic throughout interstellar space. The spectrum of the GCR consists of 98% protons and heavier ions (baryon component) and 2% electrons and positrons (lepton component). The baryon component is composed of 87% protons, 12% helium ions (alpha particles), and the remaining 1% heavy ions of charge 3 from lithium to 92 from uranium. Due to their high abundance, iron ions are highly penetrating, giving them a large potential for radiobiological damage (3,21).While the UV photochemistry of spore DNA and repair of UV damage to DNA during germination are well characterized (34,44,(59)(60)(61), there has been relatively little work on the nature of DNA damage in spores caused by ionizing radiation, the protective role of...

BackgroundThe intestinal microbial communities and their temporal dynamics are gaining increasing interest due to the significant implications for human health. Recent studies have shown the dynamic behavior of the gut microbiota in free-living, healthy persons. To date, it is not known whether these dynamics are applicable during prolonged life sharing in a confined and controlled environment.ResultsThe MARS500 project, the longest ground-based space simulation ever, provided us with a unique opportunity to trace the crew microbiota over 520 days of isolated confinement, such as that faced by astronauts in real long-term interplanetary space flights, and after returning to regular life, for a total of 2 years. According to our data, even under the strictly controlled conditions of an enclosed environment, the human gut microbiota is inherently dynamic, capable of shifting between different steady states, typically with rearrangements of autochthonous members. Notwithstanding a strong individuality in the overall gut microbiota trajectory, some key microbial components showed conserved temporal dynamics, with potential implications for the maintenance of a health-promoting, mutualistic microbiota configuration.ConclusionsSharing life in a confined habitat does not affect the resilience of the individual gut microbial ecosystem, even in the long term. However, the temporal dynamics of certain microbiota components should be monitored when programming future mission simulations and real space flights, to prevent breakdowns in the metabolic and immunological homeostasis of the crewmembers.Electronic supplementary materialThe online version of this article (doi:10.1186/s40168-017-0256-8) contains supplementary material, which is available to authorized users.

Executive Summary Return of samples from the surface of Mars has been a goal of the international Mars science community for many years. Affirmation by NASA and ESA of the importance of Mars exploration led the agencies to establish the international MSR Objectives and Samples Team (iMOST). The purpose of the team is to re‐evaluate and update the sample‐related science and engineering objectives of a Mars Sample Return (MSR) campaign. The iMOST team has also undertaken to define the measurements and the types of samples that can best address the objectives. Seven objectives have been defined for MSR, traceable through two decades of previously published international priorities. The first two objectives are further divided into sub‐objectives. Within the main part of the report, the importance to science and/or engineering of each objective is described, critical measurements that would address the objectives are specified, and the kinds of samples that would be most likely to carry key information are identified. These seven objectives provide a framework for demonstrating how the first set of returned Martian samples would impact future Martian science and exploration. They also have implications for how analogous investigations might be conducted for samples returned by future missions from other solar system bodies, especially those that may harbor biologically relevant or sensitive material, such as Ocean Worlds (Europa, Enceladus, Titan) and others. Summary of Objectives and Sub‐Objectives for MSR Identified by iMOST This objective is divided into five sub‐objectives that would apply at different landing sites. 1.1 Characterize the essential stratigraphic, sedimentologic, and facies variations of a sequence of Martian sedimentary rocks. 1.2 Understand an ancient Martian hydrothermal system through study of its mineralization products and morphological expression. 1.3 Understand the rocks and minerals representative of a deep subsurface groundwater environment. 1.4 Understand water/rock/atmosphere interactions at the Martian surface and how they have changed with time. 1.5 Determine the petrogenesis of Martian igneous rocks in time and space. This objective has three sub‐objectives: 2.1 Assess and characterize carbon, including possible organic and pre‐biotic chemistry. 2.2 Assay for the presence of biosignatures of past life at sites that hosted habitable environments and could have preserved any biosignatures. 2.3 Assess the possibility that any life forms detected are alive, or were recently alive. Summary of iMOST Findings Several specific findings were identified during the iMOST study. While they are not explicit recommendations, we suggest that they should serve as guidelines for future decision making regarding planning of potential future MSR missions. The samples to be collected by the Mars 2020 (M‐2020) rover will be of sufficient size and quality to address and solve a wide variety of scientific questions. Samples, by definition, are a statistical representation of a larger entity...