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Strukturbiologische und mechanistische Untersuchungen zur Erkennung und Reparatur von DNA-Photoschaden

Strukturbiologische und mechanistische Untersuchungen zur Erkennung und Reparatur von DNA-Photoschaden

          
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About the Book

Es ist bekannt, dass der UV-Anteil des Sonnenlichts zur Schadigung der DNA und zur Ausbildung einer Vielfalt an DNA-Photoschaden fuhrt. Die dabei entstehenden cytotoxischen und mutagenen Schaden sind Cyclobutanpyrimidindimere (CPD-Schaden), (6-4)- Photoschaden, als auch dessen Dewar Valenz-Isomere. Heutzutage besteht kein Zweifel daran, dass diese Schaden eng mit dem Auftreten von Hautkrebs in Verbindung stehen. Um sich vor den negativen Auswirkungen der Photoschaden schutzen zu konnen, haben sich alle dem Sonnenlicht ausgesetzte Lebewesen im Laufe der Evolution ein multifunktionelles und effizientes Reparatursystem angeeignet. Hier waren zum Beispiel die Exzisionsreparatur von geschadigter DNA und die direkte Reversion der Schaden zu nennen. Die letztgenannte Form der Reparatur nennt man Photoreaktivierung und wird der Enzymklasse der Photolyasen zugeschrieben. Diese Photolyasen sind paradoxerweise in der Lage, mit Hilfe von UV-A/Bbzw. Blaulicht (300 – 500 nm), Pyrimidindimere wieder in ihre intakten Basen umzuwandeln. Die Photolyasen sind hochselektive Enzyme und lassen sich je nach Substratspezifitat in CPD- und (6-4)-Photolyasen unterteilen. In den letzten Jahren hat sich der CPD-Schaden zusammen mit der CPD-Photolyase als Modellsystem bei der Untersuchung zur Entstehung und Reparatur von DNA-Photoschaden entwickelt. So sind zum Beispiel der Reparaturmechanismus sowie die Cofaktoren- Zusammensetzung der CPD-Photolyasen weitestgehend geklart. Im Gegensatz dazu weiß man vergleichsweise wenig uber die (6-4)-Schaden und ihre Photolyasen. Aufgrund der Ahnlichkeit zu den CPD-Photolyasen, wurde fur die (6-4)-Photolyasen ein ahnlicher Reparaturmechanismus postuliert, wobei jedoch der dabei auftretende viergliedrige Ubergangszustand (ein Oxetan- oder Azetidin-Intermediat) experimentell nicht nachgewiesen werden konnte. Um speziell diese als auch weitere Fragen bezuglich der (6-4)-Schadenserkennung und Reparatur durch die (6-4)-Photolyasen klaren zu konnen, musste der (6-4)-Schaden in ausreichenden Mengen fur biochemische und strukturbiologische Experimente synthetisiert werden. Im Rahmen dieser Arbeit konnte schließlich eine Methode optimiert werden, mit der man durch direkte Belichtung von Oligonukleotiden bei 254 nm unter Auschluss von Sauerstoff, ausreichende Mengen an (6-4)-Schaden enthaltener DNA isolieren konnte. Die Kristal


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Product Details
  • ISBN-13: 9783869554600
  • Binding: Paperback
  • Language: German
  • Width: 267 mm
  • ISBN-10: 3869554606
  • Height: 210 mm
  • Weight: 12 gr

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