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Probing the response of Deinococcus radiodurans exposed to simulated space conditions

Published online by Cambridge University Press:  19 November 2019

Gabriel Guarany de Araujo
Affiliation:
Interunities Graduate Program in Biotechnology, University of São Paulo, Av. Prof. Lineu Prestes, 2415, 05508-900, São Paulo, Brazil
Fabio Rodrigues
Affiliation:
Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes, 748, 05508-000, São Paulo, Brazil
Douglas Galante*
Affiliation:
Brazilian Synchrotron Light Laboratory, Brazilian Center for Research in Energy and Materials, Av. Giuseppe Máximo Scolfaro, 10000, 13083-100, Campinas, Brazil
*
Author for correspondence: Douglas Galante, E-mail: [email protected]

Abstract

The extremotolerant bacterium Deinococcus radiodurans is used as a model to explore the limits of life on Earth and beyond. In experiments performed in an ultra-high vacuum chamber with a vacuum ultraviolet (VUV) synchrotron beamline, this microorganism was exposed to conditions present on an extraterrestrial environment unprotected by an atmosphere, such as outside a spacecraft or on an asteroid, relevant in the context of planetary protection and panspermia hypothesis. Different methods were used to obtain the biologically relevant information from this investigation. Counting of colony forming units, the traditional approach for viability assessment, is limited to measuring the survival of the cells. For a more in-depth study of damage mechanisms at subcellular levels, specific molecular probes (propidium iodide and dihydrorhodamine 123) were applied and whole populations could be analysed, cell by cell, by flow cytometry. VUV radiation caused a substantial loss of viability, though only a fraction of the cells presented membrane damages even at the largest tested fluences. Additionally, intracellular oxidative stress was also detected upon exposure. These results point to significant VUV inactivating effects extending beyond the cells' outermost structures, in contrast to a more superficial role that could be expected due to the highly interacting nature of this radiation range. Nevertheless, it was observed that microscopic-level shading sufficed to allow the persistence of a small surviving subpopulation for the longer expositions. This study contributes to unveiling the response of biological systems under space conditions, assessing not just cell viability but also the mechanisms that lead to inactivation.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2019

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References

Abrevaya, XC, Paulino-Lima, IG, Galante, D, Rodrigues, F, Mauas, PJD, Corton, E and Lage, CDS (2011) Comparative survival analysis of Deinococcus radiodurans and the haloarchaea Natrialba magadii and Haloferax volcanii exposed to vacuum ultraviolet irradiation. Astrobiology 11, 10341040.CrossRefGoogle ScholarPubMed
Baatout, S, De Boever, P and Mergeay, M (2005) Temperature-induced changes in bacterial physiology as determined by flow cytometry. Annals of Microbiology 55, 7380.Google Scholar
Battista, JR (1997) Against all odds: the survival strategies of Deinococcus radiodurans. Annual Review of Microbiology 51, 203224.CrossRefGoogle ScholarPubMed
Bauermeister, A, Moeller, R, Reitz, G, Sommer, S and Rettberg, P (2011) Effect of relative humidity on Deinococcus radiodurans' resistance to prolonged desiccation, heat, ionizing, germicidal, and environmentally relevant UV radiation. Microbial Ecology 61, 715722.CrossRefGoogle ScholarPubMed
Berney, M, Weilenmann, HU and Egli, T (2006) Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS). Microbiology (Reading, England) 152, 17191729.CrossRefGoogle Scholar
Brain, DA and Jakosky, BM (1998) Atmospheric loss since the onset of the Martian geologic record: combined role of impact erosion and sputtering. Journal of Geophysical Research 103, 2268922694.Google Scholar
Cabiscol, E, Tamarit, J and Ros, J (2010) Oxidative stress in bacteria and protein damage by reactive oxygen species. International Microbiology 3, 38.Google Scholar
Cavasso Filho, RL, Homen, MGP, Fonseca, PT and Naves de Brito, A (2007) A synchrotron beamline for delivering high purity vacuum ultraviolet photons. Review of Scientific Instruments 78, 115104.CrossRefGoogle ScholarPubMed
Cockell, CS, Catling, DC, Davis, WL, Snook, K, Kepner, RL, Lee, P and McKay, CP (2000) The ultraviolet environment of Mars: biological implications past, present, and future. Icarus 146, 343359.CrossRefGoogle ScholarPubMed
Coohill, TP (1986) Virus-cell interactions as probes for vacuum ultraviolet radiation-damage and repair. Photochemistry and Photobiology 44, 359363.CrossRefGoogle ScholarPubMed
DasSarma, P, Laye, VJ, Harvey, J, Reid, C, Shultz, J, Yarborough, A, Lamb, A, Koske-Phillips, A, Herbst, A, Molina, F, Grah, O, Phillips, T and DasSarma, S (2017) Survival of halophilic Archaea in Earth's cold stratosphere. International Journal of Astrobiology 16, 321327.CrossRefGoogle Scholar
Diaz, B and Schulze-Makuch, D (2006) Microbial survival rates of Escherichia coli and Deinococcus radiodurans under low temperature, low pressure, and UV-irradiation conditions, and their relevance to possible Martian life. Astrobiology 6, 332347.CrossRefGoogle ScholarPubMed
Farci, D, Slavov, C, Tramontano, E and Piano, D (2016) The S-layer protein DR_2577 binds deinoxanthin and under desiccation conditions protects against UV-radiation in Deinococcus radiodurans. Frontiers in Microbiology 7, 155.CrossRefGoogle ScholarPubMed
Fiksdal, L and Tryland, I (1999) Effect of UV light irradiation, starvation and heat on Escherichia coli β-D-galactosidase activity and other potential viability parameters. Journal of Applied Microbiology 87, 6271.CrossRefGoogle ScholarPubMed
Fredrickson, JK, Li, SMW, Gaidamakova, EK, Matrosova, VY, Zhai, M, Sulloway, HM, Scholten, JC, Brown, MG, Balkwill, DL and Daly, MJ (2008) Protein oxidation: key to bacterial desiccation resistance? The ISME Journal 2, 393403.CrossRefGoogle ScholarPubMed
Horneck, G, Klaus, DM and Mancinelli, RL (2010) Space microbiology. Microbiology and Molecular Biology Reviews 74, 121156.CrossRefGoogle ScholarPubMed
Ito, T, Sweet, RM and Woodhead, AD (1989) Vacuum ultraviolet photobiology with synchrotron radiation. Synchrotron Radiation in Structural Biology. Boston, MA: Springer US, pp. 221241.CrossRefGoogle Scholar
Joux, F and Lebaron, P (2000) Use of fluorescent probes to assess physiological functions of bacteria at single-cell level. Microbes and Infection 2, 15231535.Google ScholarPubMed
Kawaguchi, Y, Yang, YJ, Kawashiri, N, Shiraishi, K, Takasu, M, Narumi, I, Satoh, K, Hashimoto, H, Nakagawa, K, Tanigawa, Y, Momoki, Y, Tanabe, M, Sugino, T, Takahashi, Y, Shimizu, Y, Yoshida, S, Kobayashi, K, Yokobori, S and Yamagishi, A (2013) The possible interplanetary transfer of microbes: assessing the viability of Deinococcus spp. under the ISS environmental conditions for performing exposure experiments of microbes in the Tanpopo mission. Origins of Life and Evolution of the Biosphere 43, 411428.Google ScholarPubMed
Khodadad, CL, Wong, GM, James, LM, Thakrar, PJ, Lane, MA, Catechis, JA and Smith, DJ (2017) Stratosphere conditions inactivate bacterial endospores from a mars spacecraft assembly facility. Astrobiology 17, 337350.CrossRefGoogle ScholarPubMed
Krisko, A and Radman, M (2010) Protein damage and death by radiation in Escherichia coli and Deinococcus radiodurans. Proceedings of the National Academy of Sciences of the USA 107, 1437314377.CrossRefGoogle ScholarPubMed
Mattimore, V and Battista, JR (1996) Radioresistance of Deinococcus radiodurans: functions necessary to survive ionizing radiation are also necessary to survive prolonged desiccation. Journal of Bacteriology 178, 633637.CrossRefGoogle ScholarPubMed
Nellen, J, Rettberg, P, Horneck, G, Harris, RA and Ouwehand, L (2003) A different approach to measure the cell survival of Deinococcus radiodurans. Third European Workshop on Exo-Astrobiology, vol. 545. Noordwijk, Netherlands: ESA Publications Division, pp. 249250.Google Scholar
Nicholson, WL, Munakata, N, Horneck, G, Melosh, HJ and Setlow, P (2000) Resistance of Bacillus endospores to extreme terrestrial and extraterrestrial environments. Microbiology and Molecular Biology Reviews 64, 548572.CrossRefGoogle ScholarPubMed
Olsson-Francis, K and Cockell, CS (2010) Experimental methods for studying microbial survival in extraterrestrial environments. Journal of Microbiological Methods 80, 113.CrossRefGoogle ScholarPubMed
Osman, S, Peeters, Z, La Duc, MT, Mancinelli, R, Ehrenfreund, P and Venkateswaran, K (2008) Effect of shadowing on survival of bacteria under conditions simulating the Martian atmosphere and UV radiation. Applied and Environmental Microbiology 74, 959970.CrossRefGoogle ScholarPubMed
Pashkovskaya, A, Kotova, E, Zorlu, Y, Dumoulin, F, Ahsen, V, Agapov, I and Antonenko, Y (2010) Light-triggered liposomal release: membrane permeabilization by photodynamic action. Langmuir 26, 57265733.CrossRefGoogle ScholarPubMed
Paulino-Lima, IG, Pilling, S, Janot-Pacheco, E, de Brito, AN, Barbosa, JARG, Leitao, AC and Lage, CDS (2010) Laboratory simulation of interplanetary ultraviolet radiation (broad spectrum) and its effects on Deinococcus radiodurans. Planetary and Space Science 58, 11801187.CrossRefGoogle Scholar
Pogoda de la Vega, U, Rettberg, P, Douki, T, Cadet, J and Horneck, G (2005) Sensitivity to polychromatic UV-radiation of strains of Deinococcus radiodurans differing in their DNA repair capacity. International Journal of Radiation Biology 81, 601611.Google ScholarPubMed
Pogoda de la Vega, U, Rettberg, P and Reitz, G (2007) Simulation of the environmental climate conditions on Martian surface and its effect on Deinococcus radiodurans. Advances in Space Research 40, 16721677.CrossRefGoogle Scholar
Pulschen, AA, de Araujo, GG, Carvalho, ACSR, Cerini, MF, Fonseca, LD, Galante, D and Rodrigues, F (2018) Survival of extremophilic yeasts in the stratospheric environment during balloon flights and in laboratory simulations. Applied and Environmental Microbiology 84, e01942e01918.CrossRefGoogle ScholarPubMed
Santos, AL, Oliveira, V, Baptista, I, Henriques, I, Gomes, NCM, Almeida, A, Correia, A and Cunha, A (2013) Wavelength dependence of biological damage induced by UV radiation on bacteria. Archives of Microbiology 195, 6374.CrossRefGoogle ScholarPubMed
Sarantopoulou, E, Gomoiu, I, Kollia, Z and Cefalas, AC (2011) Interplanetary survival probability of Aspergillus terreus spores under simulated solar vacuum ultraviolet irradiation. Planetary and Space Science 59, 6378.CrossRefGoogle Scholar
Sarantopoulou, E, Stefi, A, Kollia, Z, Palles, D, Petrou, PS, Bourkoula, A, Koukouvinos, G, Velentzas, AD, Kakabakos, S and Cefalas, AC (2014) Viability of Cladosporium herbarum spores under 157 nm laser and vacuum ultraviolet irradiation, low temperature (10 K) and vacuum. Journal of Applied Physics 116, 104701.CrossRefGoogle Scholar
Slade, D and Radman, M (2011) Oxidative stress resistance in Deinococcus radiodurans. Microbiology and Molecular Biology Reviews 75, 133191.CrossRefGoogle ScholarPubMed
Smith, DJ, Schuerger, AC, Davidson, MM, Pacala, SW, Bakermans, C and Onstott, TC (2009) Survivability of Psychrobacter cryohalolentis K5 under simulated Martian surface conditions. Astrobiology 9, 221228.CrossRefGoogle ScholarPubMed
Wilhelm, J, Vytasek, R, Ostadalova, I and Vajner, L (2009) Evaluation of different methods detecting intracellular generation of free radicals. Molecular and Cellular Biochemistry 328, 167176.CrossRefGoogle ScholarPubMed
Zvereva, G, Kirtsideli, I, Benken, K, Saifitdinova, A, Galkina, S, Parfenov, V, Tarasenko, VF and Kabanov, AM (2015) Investigation of the effect of VUV radiation on the viability of microfungi spores. International Conference on Atomic and Molecular Pulsed Lasers XII, vol. 9810. Tomsk, Russian Federation: SPIE, pp. 98101W.Google Scholar