Publikationsserver der Universitätsbibliothek Marburg
Titel:Molekulare Mechanismen während der Anheftung und Biofilmbildung in Shewanella oneidensis MR-1 - Die Tücken des Besiedelns -
Autor:Gödeke, Julia
Weitere Beteiligte: Bremer, Erhard (Prof. Dr.)
Veröffentlicht:2011
URI:https://archiv.ub.uni-marburg.de/diss/z2011/0466
DOI: https://doi.org/10.17192/z2011.0466
URN: urn:nbn:de:hebis:04-z2011-04668
DDC: Biowissenschaften, Biologie
Titel (trans.):Molecular Mechanisms during Attachment and Biofilm Formation in Shewanella oneidensis MR-1- Pitfalls of Colonization -
Publikationsdatum:2011-08-08
Lizenz:https://rightsstatements.org/vocab/InC-NC/1.0/

Dokument

Schlagwörter:
phage, Shewanella, Shewanella, detachment, nuclease, Bakteriophagen, Biofilm, lysis, Nuklease, Lyse <Biologie>

Zusammenfassung:
Grenzflächenbesiedlung durch robuste Bakteriengemeinschaften – sogenannten Biofilmen – stellt die ubiquitär verbreitete Lebensform von Mikroorganismen dar, um einer Vielzahl von Stressfaktoren zu widerstehen. Das Entwicklungsprogramm solcher Biofilme lässt sich in mehrere distinkte Schritte unterteilen, wobei zu Anfang eine zunächst transiente Anheftung Oberflächen- assoziierter Zellen steht, auf die eine irreversible Interaktion mit dem Substratum folgt. Nach der Bildung von Mikrokolonien entstehen im weiteren Verlauf drei-dimensionale Strukturen, die als charakteristisches Merkmal für Biofilme gelten. Das dissimilatorisch Metallionen-reduzierende Bakterium Shewanella oneidensis MR-1 bildet Biofilme unter statischen wie hydrodynamischen Bedingungen aus. Die Ausprägungsformen des Biofilms unterscheiden sich jedoch stark in Abgängigkeit von den vorherrschenden Bedingungen. So bildet S. oneidensis MR-1 unter statischen Bedingungen eine eher lockere netzartige Biofilmstruktur, durch die sich Zellen bewegen können. Hydrodynamische Bedingungen haben zur Folge, dass sich turmartige Strukturen, welche vornehmlich durch klonales Wachstum entstehen, ausbilden, die von einer dichtgepackten, widerstandsfähigen Matrix zusammengehalten werden. Um zelluläre Prozesse während der Oberflächen-Anheftung in Abhängigkeit der vorherrschenden Bedingung mit Hilfe von Transkriptom-Analysen zu identifizieren, wurde ein System zur Isolierung von Oberflächen-assoziierten Zellen in einem hydrodynamischen Umfeld etabliert. Die Transkriptom-Analysen adhärierter Zellen unter statischen und hydrodynamischen Bedingungen zeigten, dass die initiale Anheftung dieses Organismus, als erster Schritt in die Richtung eines sessilen Lebensstils im Biofilm, weitreichende Veränderungen des Transkriptoms nach sich zieht. Dabei lässt sich zwischen einer allgemeinen, jedoch umweltabhängigen Anpassung, die vor allem eine Reduzierung der Motilität und – unter statischen Bedingungen – eine rapide wie effiziente Umstellung auf anaeroben Stoffwechsel beinhaltet, und einer Substrat-spezifischen Adaption unterscheiden. So hat beispielsweise eine Anheftung an eine Eisen(hydr)oxid-Oberfläche zur Folge, dass – wahrscheinlich aufgrund des Überangebots an Elektronenakzeptoren – besonders Transportsysteme und Cytochrome reprimiert, dagegen stressabhängige Sigmafaktoren induziert werden. Im weiteren Verlauf der Anheftung und Biofilmentwicklung produzieren die S. oneidensis MR-1 Zellen eine Biofilm-Matrix, die zu einem entscheidenden Teil aus extrazellulärer DNA (eDNA) besteht. Als strukturgebendes Element spielt die eDNA in Biofilmen sowohl unter statischen, als auch unter hydrodynamischen Bedingungen eine wichtige Rolle. Die Herkunft dieser eDNA konnte auf zelllytische Prozesse – ausgelöst durch Phagen-induzierte Lyse einer Subpopulation von Zellen – zurückgeführt werden. Die durch die lytische Aktivität der drei Shewanella-eigenen Prophagen (MuSo1, MuSo2 und LambdaSo) freigesetzte DNA ist bereits in den ersten Schritten der Anheftung essenziell. Eine Mutante, in der alle drei Prophagen deletiert wurden, wies massive Defekte in allen Stadien der Biofilmbildung auf. Mutanten-Analysen und Infektionsstudien zeigten des Weiteren, dass nur die Prophagen MuSo2 und LambdaSo in den lytischen Lebenszyklus konvertieren und infektiöse Viruspartikel assemblieren können. Um die dynamische Besiedelung einer Oberfläche zu gewährleisten, gehen einige Zellen während aller Entwicklungsstufen der Biofilmbildung zu einem erneut planktonischen Lebensstil über. Dieser kontrollierte Übergang scheint unter anderem auch die Fähigkeit zur Degradierung von Komponenten der Biofilm-Matrix – insbesondere eDNA – vorauszusetzen. Zwei Kandidaten für solche DNA-degradierenden Prozesse stellen die extrazellulären Endonukleasen ExeM und ExeS dar, die vermutlich sowohl die Akkumulation, als auch das Ablösen von Biofilm-Biomasse aktiv mitgestalten, und zusätzlich – zumindest bei ExeM – in der Verwertbarkeit von DNA als Phosphatquelle eine Rolle spielen.

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