Graduation Year

2012

Document Type

Thesis

Degree

M.S.

Degree Granting Department

Biology (Cell Biology, Microbiology, Molecular Biology)

Major Professor

James T. Riordan

Keywords

ADAR, EHEC, GDAR, qRT-PCR, rpoN

Abstract

Extreme acid resistance contributes to the successful transmission of enterohemorrhagic E. coli (EHEC) through acidic food matrices and the stomach, allowing it to gain access to the intestine and elicit disease in humans. Alternative sigma factor N (σN, encoded by rpoN) was previously identified as a novel regulator of extreme acid resistance in EHEC. This study investigated the role for σN and co-expressed products of the rpoN operon in the acid resistance phenotype of EHEC. The results revealed that σN primarily controls acid resistance through repression of the glutamate-dependent acid resistance (GDAR) system through control of the σS-directed GadXW pathway. σN was also determined to repress additional acid resistance systems, including arginine-dependent acid resistance, and an anaerobic acid resistance mechanism. Two gene products of the rpoN operon, hpf and ptsN, were also determined to negatively affect GDAR, as well as expression of the σN dependent genes glnA, astA, and pspA. Mutation of hpf and ptsN did not however alter the transcription of rpoN. Transcript levels of rpoN operon genes were observed to be differential, and inconsistent with the hypothesis of expression as a single transcriptional unit. Together this data signifies the importance of rpoN operon genes in the negative regulation of extreme acid resistance systems, and suggests that the products of hpf and ptsN control the activity of σN at its promoters.

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