Characterization of different types of radiation- and pressure-induced DNA damage in Bacillus subtilis spores and their global transcriptional response during spore germination [Elektronische Ressource] / von Ralf Möller

R.Moeller

Sujets

• Biologie

Informations

• Publié par: technische_universitat_carolo-wilhelmina_zu_braunschweig
• Publié le: 01 janvier 2008
• Nombre de lectures: 43
• Langue: Deutsch
• Poids de l'ouvrage: 19 Mo

Extrait

Characterization of different types of radiation- and pressure-induced
DNA damage in Bacillus subtilis spores and their global
transcriptional response during spore germination






Von der Fakultät für Lebenswissenschaften

der Technischen Universität Carolo-Wilhelmina

zu Braunschweig

zur Erlangung des Grades eines

Doktors der Naturwissenschaften

(Dr. rer. nat.)

genehmigte

D i s s e r t a t i o n







von

Ralf Möller

aus Suhl / Deutschland



Characterization of different types of radiation- and pressure-induced
DNA damage in Bacillus subtilis spores and their global
transcriptional response during spore germination



Von der Fakultät für Lebenswissenschaften

der Technischen Universität Carolo-Wilhelmina

zu Braunschweig

zur Erlangung des Grades eines

Doktors der Naturwissenschaften

(Dr. rer. nat.)

genehmigte

D i s s e r t a t i o n













von Ralf Möller
aus Suhl





















































1. Referent: Prof. Dr. Dieter Jahn
2. Referent: Prof. Dr. Erko Stackebrandt
eingereicht am: 26.09.2007
mündliche Prüfung (Disputation) am: 18.12.2007
Druckjahr 2008

Vorveröffentlichungen der Dissertation

Teilergebnisse aus dieser Arbeit wurden mit Genehmigung der Fakultät für
Lebenswissenschaften, vertreten durch den Mentor der Arbeit, in folgenden Beiträgen vorab
veröffentlicht:

PUBLIKATIONEN

Moeller, R., G. Horneck, R. Facius, and E. Stackebrandt. 2005. Role of pigmentation in
protecting Bacillus sp. endospores against environmental UV radiation. FEMS Microbiol.
Ecol. 51:231-236.

Moeller, R., G. Horneck, P. Rettberg, H.-J. Mollenkopf, E. Stackebrandt, and W. L.
Nicholson. 2006. A method for extracting RNA from dormant and germinating Bacillus
subtilis strain 168 endospores. Curr. Microbiol. 53:227-231.

Moeller, R., G. Horneck, U. Pogoda de la Vega, P. Rettberg, E. Stackebrandt, G. Reitz, R.
Okayasu, and T. Berger. 2006. DNA damaging effects of heavy ion and X-ray irradiation of
Bacillus subtilis spores. Int. J. Astrobiol. 5:87 (abstract).

Moeller, R., E. Stackebrandt, G. Reitz, T. Berger, P. Rettberg, A. J. Doherty, G. Horneck, and
W. L. Nicholson. 2007. Role of DNA repair by non-homologous end-joining (NHEJ) in
Bacillus subtilis spore resistance to extreme dryness, mono- and polychromatic UV and
ionizing radiation. J. Bacteriol. 189:3306-3311.

Moeller, R., T. Douki, J. Cadet, E. Stackebrandt, W. L. Nicholson, P. Rettberg, G. Reitz, and
G. Horneck. 2007. UV radiation induced formation of DNA bipyrimidine photoproducts in
Bacillus subtilis endospores and their repair during germination. Int. Microbiol. 10:39-46.

Moeller, R., E. Stackebrandt, T. Douki, J. Cadet, P. Rettberg, H.-J. Mollenkopf, G. Reitz, and
G. Horneck. 2007. DNA bipyrimidine photoproduct repair and transcriptional response of
UV-C irradiated Bacillus subtilis. Arch. Microbiol. (Epub ahead of print;
DOI: 10.1007/s00203-007-0263-4).

Stöffler, D., G. Horneck, S. Ott, U. Hornemann, C. S. Cockell, R. Moeller, C. Meyer, J. P. de
Vera, J. Fritz, and N. A. Artemieva. 2007. Experimental evidence for the potential impact
ejection of viable microorganisms from Mars and Mars-like planets. Icarus 186:585-588.

Edwards, H. G. M., R. Moeller, S. E. Jorge Villar, G. Horneck, and E. Stackebrandt. 2006.
Raman spectroscopic study of the photoprotection of extremophilic microbes against
ultraviolet radiation. Int. J. Astrobiol. 5:313-318.

Wassmann, M., R. Moeller, J. Nellen, G. Reitz, E. Rabbow, and P. Rettberg. 2007. Bacillus
subtilis’ ability to adapt to extreme UV stress. Int. J. Astrobiol. 6:71 (abstract).

Horneck, G., D. Stöffler, S. Ott, U. Hornemann, C. S. Cockell, R. Moeller, C. Meyer, J. P. de
Vera, J. Fritz, S. Schade, and N. A. Artemieva. 2007. Microbial Rock Inhabitants Survive
Hypervelocity Impacts on Mars-like Host Planets: First Phase of Lithopanspermia
Experimentally Tested. Astrobiology (AST-2007-0134.R1; accepted for publication).



TAGUNGSBEITRÄGE

Moeller, R., G. Horneck, E. Stackebrandt, H. G. M. Edwards, and S. E. Jorge Villar. 2003.
rdDo endogenous pigments protect Bacillus spores against UV-radiation? 3 European
Workshop on Exo-/Astrobiology, Mars: The search for Life, ESA SP-545:241-243 (abstract).

Moeller, R., E. Stackebrandt, H. G. M. Edwards, and G. Horneck. 2004. Bacterial endospores
stretch the limits of survivability. EMBO conference on Molecular Microbiology, Heidelberg,
Germany, Topic: Exploring prokaryotic diversity, p. 97 (abstract).

Moeller, R., and G. Horneck. 2004. Bacillus endospores - an ideal exobiological tool.
thAbstracts of the 35 Committee on Space Research (COSPAR) Scientific Assembly, Paris,
France (COSPAR04-A-02596) (abstract).

Moeller, R., G. Horneck, E. Stackebrandt, G. Reitz, P. Rettberg, U. Pogoda de la Vega, R.
Okayasu, and T. Berger. 2005. DNA damaging effects of heavy ion and X-ray irradiation of
thBacillus subtilis spores. 5 European Workshop on Exo-/Astrobiology, Budapest, Hungary
(poster).

Moeller, R., T. Berger, G. Reitz, and R. Okayasu. 2006. Gene activation of heavy ion treated
Bacillus subtilis 168 (DSM 402) endospores during germination involved DNA-repair - first
results. Annual Report of the Research Project with Heavy Ions at NIRS-HIMAC 2005/2006
(abstract).

Moeller, R., G. Horneck, P. Rettberg, and E. Stackebrandt. 2006. Induced DNA damage by
simulated space conditions in Bacillus subtilis endospores. DFG workshop “Viability in
Space” Abstract book, p. 52-53, Cologne, Germany (abstract).

Moeller, R., G. Horneck, P. Rettberg, and E. Stackebrandt. 2006. Gene activation of Bacillus
subtilis after UV irradiation”, BIOspektrum (special edition, 2006), p. 77, Annual VAAM
Meeting, Jena, Germany (abstract).

Moeller, R., T. Berger, G. Reitz, M. Okada, and R. Okayasu. 2006 and 2007. Gene activation
of heavy ion treated Bacillus subtilis 168 endospores during germination involved
DNArepair (17B-463). HIMAC Heavy Ion Researcher Investigator Meeting at NIRS, Inage, Japan
(poster).

Moeller, R., G. Horneck, D. Stöffler, S. Ott, U. Hornemann, C. Meyer, J. P. de Vera, and J.
Fritz. 2006. Could microorganisms survive an impact ejection? Deutsche Physikalische
Gesellschaft - Annual meeting in Heidelberg, Germany (abstract).

Moeller, R., T. Berger, G. Reitz, and R. Okayasu. 2007. Gene activation of heavy ion treated
Bacillus subtilis 168 (DSM 402) endospores during germination involved DNA-repair - site
specific mutagenesis and oxidative DNA damage induction. Annual Report of the Research
Project with Heavy Ions at NIRS-HIMAC 2006/2007 (abstract).

Moeller, R., E. Stackebrandt, G. Horneck, T. Berger, P. Rettberg, R. Okayasu, and G. Reitz.
2007. NHEJ - a DNA strand breaks mechanism of Bacillus subtilis spores. BIOspektrum
(special edition, 2007), p. 83; Annual VAAM Meeting, Osnabrück, Germany (abstract).













“All truths are easy to understand once they are discovered; the point is to discover them.”
(Galileo Galilei, 1564 - 1642)









This thesis is dedicate to my mother
(Marion Möller, 1949-2006).

TABLE OF CONTENTS
INTRODUCTION 1
1. Introduction 1
1.1 Bacterial endospores stretch the limits of life 1
1.2 Impact of sporulation on the life cycle of Bacillus subtilis 2
1.3 Factors contributing to spore resistance 5
1.4 Role of germination in spore’s survival 10
1.5 DNA repair systems 12
1.6 Aim of this thesis 19

MATERIAL AND METHODS 21
2. Material and methods 21
2.1 Cultivation of Bacillus subtilis and nucleic acids isolation 21
2.1.1 Media 21
2.1.2 Bacillus subtilis strains 22
2.1.3 Cultivation of vegetative cells 23
2.1.4 Sporulation, spore purification and storage 23
2.1.5 Assay for colony formation ability 23
2.1.6 Spore germination 24
2.1.7 DNA extraction 24
2.1.8 RNA isolation and quantification 25
2.2 Experimental conditions and exposure facilities 26
2.2.1 Experimental strategy for studying DNA repair processes in B. subtilis spores 27
2.2.2 Sample preparation 27
2.2.2.1 Air-dried spore (mono-)layers and sample recovery via “PVA-stripping” 27
2.2.2.2 Spores in aqueous suspension 28
2.2.3 UV irradiation 29
2.2.3.1 UV-C irradiation of B. subtilis spores 29
2.2.3.2 Polychromatic UV