Date of Award

12-15-2006

Degree Type

Thesis

Degree Name

M.S.

Degree Program

Biological Sciences

Department

Biological Sciences

Major Professor

Clancy, Mary

Second Advisor

Timpte, Candace

Third Advisor

Hurlburt, Barry

Abstract

Spx is a global regulator discovered in Bacillus subtilis to suppress basal growth and development processes and activate transcription of genes involved in thiol homeostasis when a cell encounters oxidative stress. Its activity relies on reversible thiol-disulfide bond formation and binding RNA polymerase rather than DNA. The discovery that Staphylococcus aureus global virulence regulator, SarA, is more active upon cysteine reduction suggests that redox response could mediate virulence in this important human pathogen. We describe the cloning of spx from S. aureus strain RN6390, overexpression in Escherichia coli, and purification of native protein. Antibodies against Spx were raised for western analysis. Spx from S. aureus was highly active in a B. subtilis in vitro transcription system, stimulating expression of trxB, the gene encoding thioredoxin reductase, without reducing agents. RNA polymerase was partially purified from S. aureus, and the enzyme was active, catalyzing transcription of rpsD, but not trxB.

Rights

The University of New Orleans and its agents retain the non-exclusive license to archive and make accessible this dissertation or thesis in whole or in part in all forms of media, now or hereafter known. The author retains all other ownership rights to the copyright of the thesis or dissertation.

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