Microbiology

Reactions of the DNA translocase FtsK and XerC & XerD recombinases from Pseudomonas aeruginosa

Luhai Xu
Student number 3107041
Supervisor Ian Grainge

Table of Contents

Abstract 3

LIST OF ABBREVIATIONS 4

Chapter1: Introduction 5

Chapter 2 Materials and Methods 26

Chapter 3: Results 41

Chapter 4: Discussion 63

References 74

Appendix 80

Abstract
DNA replication is an important biological process and occurs in all living organisms to copy their DNA. However, chromosome dimers might be formed by homologous recombination between sister chromosome before cell division which is lethal to the cell. Luckily, these chromosome dimers can be resolved back into monomers by a site-specific recombination carried out at the dif site prior to cell divison.
XerC/XerD recombination is an example of site-specific recombination found in almost all bacteria. Three critical factors are involved in this recombination reaction: the DNA translocase FtsK, tyrosine recombinases XerC/XerD and the specific site dif. The chromosome dimer resolution is done after the activation of XerD by the γ sub-domain of FtsK at the dif site. When stimulated by FtsK, XerD exchanges the first pair of strands to form a Holliday junction which is then resolved by XerC.
FtsK is a multidomain protein required for cell division and chromosome partitioning. It consists of an N-terminal domain, linker domain and C-terminal domain. The C-terminal domain is made up of three sub-domains α，β and γ. Its multidomain makes it is multifunctional. FtsK acts at the bacterial division septum to couple chromosome segregation with cell division. It also promotes the XerC/XerD site-specific recombination at dif sites by switching the activity of the XerC/XerD recombinases. XerC and XerD, two recombinases belonging to the lambda integrase family of enzymes, are essential for the Xer site-specific recombination.
This project concerns work on the mechanisms

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