Date of Award

2009

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

Dr. David Haniford

Second Advisor

Dr. David Edgell

Third Advisor

Dr. Gary Shaw

Abstract

Host factor proteins, IHF and H-NS, regulate TnlO transposition by mediating transpososome assembly and by providing stability to the transpososome to perform transposition. To investigate the structural role on the TnlO transpososome provided by H-NS, relative stabilities of the folded and unfolded transpososome forms bound to H-NS were determined. H-NS was found to bind stronger to the unfolded transpososome in promoting intermolecular transposition. The impact of an increased IHF affinity on transposition was explored with a transpososome that resisted unfolding in order to examine the relationship between transpososome unfolding and transposon excision. The unfolding defect led to impaired transposon excision, hence affecting strand transfer efficiency. Strand transfer efficiency was improved by adding H-NS, although excision remained defective. Together, IHF and H-NS regulate TnlO transposition by modulating structural transitions in the transpososome to promote propagation of the transposon and to enhance fitness of the transposon for evolutionary purposes

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