Date of Award

2006

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

Dr. David Haniford

Second Advisor

Dr Megan Davey

Third Advisor

Dr. David Litchfield

Abstract

H-NS enhances TnlO transposition by directly binding to the transposition complex (transpososome) and unfolding it. In an unfolded state, inteπnolecular transposition events are favored. The purpose of this study was to further understand what functions of H-NS are required for the formation of a stably unfolded transpososome. To determine what functions of H-NS are essential, specific H-NS mutants were generated and tested for their abilities to stimulate transposition in vitro and interact with the transpososome. Four major conclusions regarding H-NS interaction with the Tnlθ transpososome were drawn. First, H-NS initially binds the transpososome through DNA structure specific recognition. Secondly, upon unfolding of the transpososome, H-NS makes a direct protein-protein interaction with transposase. Thirdly, H-NS alters the IHF binding site to potentially prevent IHF from refolding the transpososome. Finally, H-NS is able to influence target site selection in TnlO transposition.

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