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J Bacteriol, February 1998, p. 705-713, Vol. 180, No. 3
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Regulation of carAB Expression in
Escherichia coli Occurs in Part through UTP-Sensitive
Reiterative Transcription
Xiaosi
Han and
Charles L.
Turnbough Jr.*
Department of Microbiology, University of
Alabama at Birmingham, Birmingham, Alabama 35294
Received 19 September 1997/Accepted 24 November 1997
In Escherichia coli, expression of the
carAB operon is subject to cumulative repression, which
occurs by ArgR-mediated repression at a downstream promoter, P2, and by
pyrimidine-mediated regulation at an upstream promoter, P1. In this
study, we show that pyrimidine-mediated regulation occurs in part
through a mechanism involving UTP-sensitive reiterative transcription
(i.e., repetitive addition of U residues to the 3' end of a nascent
transcript due to transcript-template slippage). In this case,
reiterative transcription occurs at the end of a run of three T
· A base pairs in the initially transcribed region of the
carAB P1 promoter. The sequence of this region is 5'-GTTTGC (nontemplate strand). In the proposed regulatory
mechanism, increased intracellular levels of UTP promote reiterative
transcription, which results in the synthesis of transcripts with the
sequence GUUUUn (where n = 1 to >30). These transcripts are not extended downstream to include
structural gene sequences. In contrast, lower levels of UTP enhance
normal template-directed addition of a G residue at position 5 of the
nascent transcript. This addition precludes reiterative transcription
and permits normal transcript elongation capable of producing
translatable carAB transcripts. Thus, carAB
expression, which is necessary for pyrimidine nucleotide (and arginine)
biosynthesis, increases in proportion to the cellular need for UTP. The
proposed mechanism appears to function independently of a second
pyrimidine-mediated control mechanism that involves the regulatory
proteins CarP and integration host factor.
*
Corresponding author. Mailing address: Department of
Microbiology, 409 BBRB/Box 6, University of Alabama at Birmingham,
Birmingham, AL 35294. Phone: (205) 934-6289. Fax: (205) 975-5479. E-mail: chuckt{at}uab.edu.
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