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A more recent version of this article appeared on October 5, 2001
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Papers In Press, published online ahead of print July 31, 2001
J. Biol. Chem, 10.1074/jbc.M104764200
Submitted on May 24, 2001
Revised on July 30, 2001
Accepted on July 30, 2001

The anti-ITS, a portable sequence that impedes promoter escape, requires sigma 70 for function

Cathleen L. Chan and Carol A. Gross

Stomatology, and Immunology and Microbiology, University of California, San Francisco, San Francisco, CA 94143

Corresponding Author: cgross{at}cgl.ucsf.edu

The anti sequence, a portable element extending from position +1 to +15 of the transcript is sufficient to prevent promoter escape from a variety of strong sigma 70 promoters. We show here that this sequence does not function with even the strongest sigma 32 promoter. Moreover, a particular class of substitutions in sigma 70 that disrupt interaction between Region 2.2 of sigma 70 and a coiled-coiled motif in the {b}’ subunit of RNA polymerase, antagonizes the function of the anti element. This same group of mutants prevents lQ-mediated anti-termination at the {l}PR’ promoter. At this promoter, interaction of sigma 70 with the non-template strand of the initial transcribed sequence (ITS) is required to promote the pause prerequisite for anti-termination. These mutants prevent pausing because they are defective in this recognition event. By analogy, we suggest that interaction of sigma 70 with the non-template strand of the anti-ITS is required for function of this portable element, thus explaining why neither sigma 32 nor the Region 2.2 sigma 70 mutants mediate anti function. Support for the analogy with the {l}PR’ promoter comes from preliminary experiments suggesting that the anti-ITS, like the {l}PR’ ITS, is bipartite.


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