Project description:Brines, potentially formed by the deliquescence and freezing point depression of highly hygroscopic salts, such as perchlorates (ClO4-), may allow for the spatial and temporal stability of liquid water on present-day Mars. It is therefore of great interest to explore the microbial habitability of Martian brines, for which our current understanding is, however, still limited. Putative microbes growing in the perchlorate-rich Martian regolith may be harmed due to the induction of various stressors including osmotic, chaotropic, and oxidative stress. We adapted the model organism Escherichia coli to increasing sodium perchlorate concentrations and used a proteomic approach to identify the adaptive phenotype. Separately, the microbe was adapted to elevated concentrations of NaCl and glycerol, which enabled us to distinguish perchlorate-specific adaptation mechanisms from those in response to osmotic, ion and water activity stress. We found that the perchlorate-specific stress-response focused on pathways alleviating damage to nucleic acids, presumably caused by increased chaotropic and/ or oxidative stress. The significant enrichments that have been found include DNA repair, RNA methylation and de novo IMP biosynthesis. Our study provides insights into the adaptive mechanisms necessary for microorganisms to survive under perchlorate stress, with implications for understanding the habitability of Martian brines.
Project description:We designed an experimental setup to investigate the transcriptomic and proteomic responses of the hyperthermophilic archaeon Pyrococcus furiosus to heat and cold shock. P. furiosus is a model organism for studying microbial adaptation to extreme environments, including deep-sea hydrothermal vents with temperature gradients ranging from 1°C to 400°C. We aimed to simulate critical conditions where P. furiosus cannot grow and to examine the immediate response to thermal stress as well as the recovery process.
Project description:Because of their ubiquity and resistance to spacecraft decontamination, bacterial spores are considered likely potential forward contaminants on robotic missions to Mars. Thus it is important to understand their global responses to long-term exposure to space or Mars environments. As part of the PROTECT experiment, spores of B. subtilis 168 were exposed to real space conditions and to simulated martian conditions for 559 days in low Earth orbit mounted on the EXPOSE-E exposure platform outside the European Columbus module on the International Space Station. Upon return, spores were germinated, total RNA extracted and fluorescently labeled, and used to probe a custom Bacillus subtilis microarray to identify genes preferentially activated or repressed relative to ground control spores. Increased transcript levels were detected for a number of stress-related regulons responding to DNA damage (SOS response, SPβ prophage induction), protein damage (CtsR/Clp system), oxidative stress (PerR regulon) and cell envelope stress (SigV regulon). Spores exposed to space demonstrated a much broader and more severe stress response than spores exposed to simulated Mars conditions. The results are discussed in the context of planetary protection for a hypothetical journey of potential forward contaminant spores from Earth to Mars and their subsequent residence on Mars. Two-color microarrays were performed comparing germination of Space-exposed or Mars-exposed vs. ground-control (Earth) spores.
Project description:At hydrothermal vent sites, chimneys consisting of sulfides, sulfates, and oxides are formed upon contact of reduced hydrothermal fluids with oxygenated seawater. The walls and surfaces of these chimneys are an important habitat for vent-associated microorganisms. We used community proteogenomics to investigate and compare the composition and in situ protein expression of microbial communities colonizing two actively venting hydrothermal chimneys from the Manus Basin back-arc spreading center (Papua New Guinea).