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J. Biol. Chem., Vol. 278, Issue 43, 41691-41701, October 24, 2003

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Poly(A)-dependent Transcription Termination

CONTINUED COMMUNICATION OF THE POLY(A) SIGNAL WITH THE POLYMERASE IS REQUIRED LONG AFTER EXTRUSION IN VIVO^*

Steven J. Kim and Harold G. Martinson

From the Department of Chemistry and Biochemistry and the Molecular Biology Institute, 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. An unsolved problem is how the poly(A) signal triggers the polymerase to terminate. A popular model is that this occurs during extrusion of the poly(A) signal, at which time it interacts with factors on the transcription complex. To test this idea we used cis-antisense inhibition in vivo to probe the temporal relationship between poly(A) signal extrusion and the commitment of the polymerase to terminate. Our rationale was to inactivate the poly(A) signal at increasing times post-extrusion to determine the point beyond which it is no longer required for termination. We found that communication with the polymerase is not temporally restricted to the time of poly(A) signal extrusion, but is ongoing and perhaps random. Some polymerases terminate almost immediately. Others have yet to receive their termination instructions from the poly(A) signal even 500 bp downstream, as indicated by the ability of an antisense at this distance to block termination. Thus, the poly(A) signal can functionally interact with the polymerase at considerable distances down the template. This is consistent with the emerging picture of a processing apparatus that assembles and matures while riding with the polymerase.

Received for publication, June 16, 2003 , and in revised form, July 10, 2003.

^* 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. This 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{at}chem.ucla.edu.

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