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J. Biol. Chem., Vol. 277, Issue 45, 42899-42911, November 8, 2002
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From the Department of Chemistry and Biochemistry, University of
California at Los Angeles, Los Angeles, California 90095-1569
Genes encoding polyadenylated mRNAs depend on
their poly(A) signals for termination of transcription. Typically,
transcription downstream of the poly(A) signal gradually declines to
zero, but often there is a transient increase in polymerase density
immediately preceding the decline. Special elements called pause sites
are traditionally invoked to account for this increase. Using run-on transcription from the nuclei of transfected cells, we show that both
the pause and the gradual decline that follow a poly(A) site are
generated entirely by the poly(A) signal itself in a series of model
constructs. We found no other elements to be involved and argue that
the elements called pause sites do not function through pausing. Both
the poly(A)-dependent pause and the subsequent decline
occurred earlier for a stronger poly(A) signal than for a weaker one.
Because the gradual decline resembles the abortive elongation that
occurs downstream of many promoters, one model has proposed that the
poly(A) signal flips the polymerase from the elongation mode to the
abortive mode like a binary switch. We compared abortive elongators
with poly(A) terminators and found a 4-fold difference in processivity.
We conclude that poly(A) terminating polymerases do not merely
revert to their prior state of low processivity but rather convert to a
new termination-prone condition.
The Poly(A) Signal, without the Assistance of Any Downstream
Element, Directs RNA Polymerase II to Pause in
Vivo and Then to Release Stochastically from the
Template*
*
This work was supported by National Institutes of Health
Grant GM50863.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. Tel.: 310-825-3767;
Fax: 310-206-4038; E-mail: hgm@chem.ucla.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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