Regulation of papBA transcription by the leucine-responsive regulatory and catabolite activator proteins.

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Title Regulation of papBA transcription by the leucine-responsive regulatory and catabolite activator proteins.
Publication Type dissertation
School or College School of Medicine
Department Pathology
Author Weyand, Nathan J.
Date 2000-08
Description The pyelonephritis-associated pili (pap) operon in Escherichia coli is regulated by an epigenetic mechanism involving the formation of specific DNA methylation patterns characteristic of transcriptionally active (phase ON) and inactive (phase OFF) cells. The binding site for cyclic AMP receptor protein (CAP) regulates the switch between phase ON and phase OFF states. Activating region 1 of CAP was shown to activate papBA transcription by a Class I-like mechanism from a distance -215.5) greater than any other that has been found in E coli. CAP does not act as an antirepressor, as was previously suggested. Instead, CAP plays an active role in stimulating papBA transcription, possibly by interacting with the C-terminal domain of the alpha subunit of RNA polymerase. An in vitro transcription system and methylation protection assay were developed using supercolied plasmid DNA to study the phase OFF (repression) and phase ON (activation) transcription states of papBA. Binding of Lrp to a Dam target site proximal to the papBA promoter (designated GATC[prox]) established a DNA methylation pattern and transcription state identical to that observed in vivo in phase OFF cells. Two pap regulatory mutations were used (pap-13 and pap-1) to study the phase ON state of pap transcription because they are constitutively (locked ON) transcribed in vivo. Binding of Lrp to a Dam target site distal to the papBA promoter (designated GATC[dis]) blocked methylation of this site and correlated with activation of transcription. Lrp is sufficient to activate transcription from pap-13 but not pap-1 template. CAP further stimulates pap-13 template transcription in an Lrp dependent manner. This thesis establishes an in vitro transcription system that has allowed the study of repression and activation of pap transcription by two global regulators of E. coli gene regulation, CAP and Lrp.
Type Text
Publisher University of Utah
Subject Escherichia coli; DNA
Subject MESH Pyelonephritis; Genetics
Dissertation Institution University of Utah
Dissertation Name PhD
Language eng
Relation is Version of Digital reproduction of "Regulation of papBA transcription by the leucine-responsive regulatory and catabolite activator proteins." Spencer S. Eccles Health Sciences Library. Print version of "Regulation of papBA transcription by the leucine-responsive regulatory and catabolite activator proteins." available at J. Willard Marriott Library Special Collection. QR6.5 2000 .W49.
Rights Management © Nathan J. Weyand.
Format application/pdf
Format Medium application/pdf
Identifier us-etd2,3963
Source Original: University of Utah Spencer S. Eccles Health Sciences Library (no longer available).
ARK ark:/87278/s6fn1mtp
Setname ir_etd
ID 193133
Reference URL https://collections.lib.utah.edu/ark:/87278/s6fn1mtp
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