DARWIN Digitale Dissertationen German Version Strich

FU Berlin
Digitale Dissertation

Jan Deneke :
The tyrosineintegrase-analog TelN catalyses the telomere resolution in the bacteriophage N15
Das Tyrosinintegrase-Analog TelN katalysiert die telomere resolution im Bakteriophagen N15

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Abstract

In this study, an in vitro system for the biogenesis of linear plasmids with hairpin ends was developed for the first time. Using the E. coli bacteriophage N15 as a model-system, it was shown, that two phage-encoded components are sufficient for the reaction. These are the protelomerase TelN and the corresponding DNA-substrate. The described reaction converts circular into linear DNA with covalently closed ends. The substrate for the reaction is a double-stranded, palindromic DNA-sequence with a length of 56 bp. The central part of the telomere resolution site (22 bp) is essential for substrate recognition and generation of the hairpin ends. The peripheric parts of the palindrome effect the high sequence-specific DNA-binding ability of TelN to its substrate. TelN is a highly specialized integrase, for which the donor-DNA is also the acceptor-DNA. During the cleaving-joining reaction the DNA is first cleaved sequence-specifically, and TelN-DNA intermediates are generated, in which the protein binds covalently to the 3?-end of each strand of the resolution site. In a second step the phosphodiester bond with the 5?-end of the complementing DNA strand is created. It is possible, that the DNA has Z-DNA conformation during the reaction. Results, suggesting this hypothesis are discussed. The in vitro-system, that is described here served as a model for the telomere resolution in Borellia burgdorferi, a human pathogen and the causing agent of Lyme disease. Similarities and dissimilarities between this and related systems are discussed.

Table of Contents

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Titelblatt und Inhalt
1. Einleitung 1
2. Material 11
3. Methoden 30
4. Ergebnisse 42
5. Diskussion 76
6.7. Summary u. Zusammenfassung 90
8.9. Abkürzungen u. Literatur 92

More Information:

Online available: http://www.diss.fu-berlin.de/2002/224/indexe.html
Language of PhDThesis: german
Keywords: telomere resolution phage N15 tyrosine integrase replication prophage TelN proteine covalently closed DNA
DNB-Sachgruppe: 30 Chemie
Date of disputation: 29-Oct-2002
PhDThesis from: Fachbereich Biologie, Chemie, Pharmazie, Freie Universität Berlin
First Referee: Prof. Dr. Walter Messer
Second Referee: Prof. Dr. Ralf Erdmann
Contact (Author): deneke@molgix.molgen.mpg.de
Contact (Advisor): messer@molgen.mpg.de
Notes: Acrobat 5.0 erforderlich
Date created:31-Oct-2002
Date available:01-Nov-2002

 

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