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J. Biol. Chem., Vol. 276, Issue 23, 19836-19844, June 8, 2001
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From the The momP1 promoter of the
bacteriophage Mu mom operon is an example of a weak
promoter. It contains a 19-base pair suboptimal spacer between
the
Intrinsic DNA Distortion of the Bacteriophage Mu
momP1 Promoter Is a Negative Regulator of Its
Transcription
A NOVEL MODE OF REGULATION OF TOXIC GENE EXPRESSION*
§,
Department of Microbiology and Cell Biology,
Indian Institute of Science, Bangalore 560 012, India and the
¶ Department of Biology, University of Rochester,
Rochester, New York 14627
35 (ACCACA) and 10 (TAGAAT) hexamers. Escherichia coli RNA polymerase is unable to bind to momP1 on its
own. DNA distortion caused by the presence of a run of six T
nucleotides overlapping the 5' end of the 10 element might prevent
RNA polymerase from binding to momP1. To investigate the
influence of the T6 run on momP1 expression,
defined substitution mutations were introduced by site-directed
mutagenesis. In vitro probing experiments with copper
phenanthroline ((OP)2Cu) and DNase I revealed distinct differences in cleavage patterns among the various mutants; in addition, compared with the wild type, the mutants showed an increase (variable) in momP1 promoter activity in vivo.
Promoter strength analyses were in agreement with the ability of these
mutants to form open complexes as well as to produce
momP1-specific transcripts. No significant role is
attributed to the overlapping and divergently organized promoter,
momP2, in the expression of momP1 activity, as
determined by promoter disruption analysis. These data support the view
that an intrinsic DNA distortion in the spacer region of
momP1 acts in cis as a negative element in
mom operon transcription. This is a novel mechanism of
regulation of toxic gene expression.
*
This work was supported by a grant from the Department of
Science and Technology, Government of India (to V. N.) and by
United States Public Health Service Grant GM29227 from the National
Institutes of Health (to S. H.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Microbiology and Cell Biology, Indian Inst. of Science, Bangalore 560 012, India. Tel: 80-360-0668 or 80-309-2598; Fax: 80-360-2697; E-mail: vraj@mcbl.iisc.ernet.in.
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