Molecular architecture and function of adenovirus DNA polymerase

Abstract

Central to this thesis is the role of adenovirus DNA polymerase (Ad pol) in adenovirus DNA replication. Ad pol is a member of the family B DNA polymerases but belongs to a distinct subclass of polymerases that use a protein as primer. As Ad pol catalyses both the initiation and elongation phases and needs to accomodate both DNA and protein as a primer, it is not surprising that a large number of protein-protein and protein-DNA interactions are involved in efficient replication. Indeed, Ad pol is known to interact with pTP, NFI and DNA, although our understanding of these interactions is limited. In this thesis, these interactions have been studied in greater detail. After an introductory chapter on DNA dependent DNA polymerases and Ad replication, the jumping back mechanism that characterizes the change from initiation to elongation is extensively reviewed in chapter 2. In chapter 3, the highly conserved (I/Y)XGG motif of Ad pol is studied. In chapter 4, the interaction between Ad pol and DNA is further studied by the use of biotinylated oligo-nucleotides with a bulky streptavidin block. Chapter 5 examines the termination of Ad pol on the native TP-containing viral DNA. Finally, in chapter 6 the recruitment of the pTP-pol complex via a direct interaction between Ad pol and NFI is studied in detail.