Dissertation/Thesis Abstract

Crystallographic Studies of the E. coli DNA Replication Restart Primosome
by Izaac, Aude E., M.S., The University of Toledo, 2005, 201; 10835921
Abstract (Summary)

Understanding the mechanisms of DNA replication has been one of the main research topics in biochemistry for decades, due to the tremendous potential applications involved, such as medical treatments and better knowledge of the biological evolution of organisms. In this perspective, macromolecular X ray crystallography provides a very powerful tool. By solving the 3 dimensional structure of the proteins involved in replication mechanisms, one can get a good insight at complicated biological systems at the atomic level. It is also important to study the protein-protein and protein-substrate interactions when they form biological complexes. This work focused on studying several proteins which are associated with the assembly of the replication restart primosome in Escherichia coli, during the repair of damaged DNA. The crystallization of PriA, the most important of the restart primosome proteins, was investigated. Two truncations of PriA (the PriA N and PriA NI domains) were expressed, purified and characterized to be prepared for crystallization. Three additional replication restart proteins: PriB, PriC and DnaT, were expressed and purified. The solubility of each protein was optimized by identifying the best solution conditions. The proteins were characterized, alone and in complexes, using several biophysical techniques. DnaT was successfully crystallized and screened for X-ray diffraction. Finally, using ten standard test proteins, the correlation between the optimization of protein solubility (and solution conditions) and the success of crystal screening was studied.

Indexing (document details)
Advisor: Mueser, Timothy
School: The University of Toledo
Department: Chemistry
School Location: United States -- Ohio
Source: MAI 57/05M(E), Masters Abstracts International
Subjects: Chemistry
Keywords: Primosome, Protein crystallography, Solubility
Publication Number: 10835921
ISBN: 978-0-355-97063-0
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