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Originally published In Press as doi:10.1074/jbc.M003149200 on July 5, 2000

J. Biol. Chem., Vol. 275, Issue 38, 29800-29807, September 22, 2000
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The Brf and TATA-binding Protein Subunits of the RNA Polymerase III Transcription Factor IIIB Mediate Position-specific Integration of the Gypsy-like Element, Ty3*

Lynn YiehDagger §, George Kassavetis, E. Peter Geiduschek, and Suzanne B. SandmeyerDagger ||**

From the Dagger  Department of Microbiology and Molecular Genetics and || Department of Biological Chemistry, University of California, Irvine, California 92697-1700 and the  Department of Biology and Center for Molecular Genetics, University of California, San Diego, La Jolla, California 92093

Ty3 integrates into the transcription initiation sites of genes transcribed by RNA polymerase III. It is known that transcription factors (TF) IIIB and IIIC are important for recruiting Ty3 to its sites of integration upstream of tRNA genes, but that RNA polymerase III is not required. In order to investigate the respective roles of TFIIIB and TFIIIC, we have developed an in vitro integration assay in which Ty3 is targeted to the U6 small nuclear RNA gene, SNR6. Because TFIIIB can bind to the TATA box upstream of the U6 gene through contacts mediated by TATA-binding protein (TBP), TFIIIC is dispensable for in vitro transcription. Thus, this system offers an opportunity to test the role of TFIIIB independent of a requirement of TFIIIC. We demonstrate that the recombinant Brf and TBP subunits of TFIIIB, which interact over the SNR6 TATA box, direct integration at the SNR6 transcription initiation site in the absence of detectable TFIIIC or TFIIIB subunit B". These findings suggest that the minimal requirements for pol III transcription and Ty3 integration are very similar.

* This work was supported in part by United States Public Health Service Grants GM33281 (to the University of California, Irvine) and GM18386 (to the University of California, San Diego) from the National Institutes of Health.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.

§ Supported by a University of California Biotechnology Research and Education Program Training Grant.

** To whom correspondence should be addressed. E-mail: sbsandme@uci.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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