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Originally published In Press as doi:10.1074/jbc.M101273200 on June 28, 2001

J. Biol. Chem., Vol. 276, Issue 34, 31786-31792, August 24, 2001
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The Drosophila U1 and U6 Gene Proximal Sequence Elements Act as Important Determinants of the RNA Polymerase Specificity of Small Nuclear RNA Gene Promoters in Vitro and in Vivo*

Kathleen J. McNamara-Schroeder, Roger F. Hennessey, Gale A. Harding, Richard C. JensenDagger , and William E. Stumph§

From the Department of Chemistry and Molecular Biology Institute, San Diego State University, San Diego, California 92182-1030

Transcription of genes coding for metazoan spliceosomal snRNAs by RNA polymerase II (U1, U2, U4, U5) or RNA polymerase III (U6) is dependent upon a unique, positionally conserved regulatory element referred to as the proximal sequence element (PSE). Previous studies in the organism Drosophila melanogaster indicated that as few as three nucleotide differences in the sequences of the U1 and U6 PSEs can play a decisive role in recruiting the different RNA polymerases to transcribe the U1 and U6 snRNA genes in vitro. Those studies utilized constructs that contained only the minimal promoter elements of the U1 and U6 genes in an artificial context. To overcome the limitations of those earlier studies, we have now performed experiments that demonstrate that the Drosophila U1 and U6 PSEs have functionally distinct properties even in the environment of the natural U1 and U6 gene 5'-flanking DNAs. Moreover, assays in cells and in transgenic flies indicate that expression of genes from promoters that contain the "incorrect" PSE is suppressed in vivo. The Drosophila U6 PSE is incapable of recruiting RNA polymerase II to initiate transcription from the U1 promoter region, and the U1 PSE is unable to recruit RNA polymerase III to transcribe the U6 gene.


* This work was supported by National Science Foundation Grant MCB-9818000 and in part by the California Metabolic Research Foundation.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.

Dagger Present address: Dept. of Immunology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, CA 92037.

§ To whom correspondence should be addressed: Dept. of Chemistry, San Diego State University, 5500 Campanile Dr., San Diego, CA 92182-1030. Tel.: 619-594-5575; Fax: 619-594-4634; E-mail: wstumph@sciences.sdsu.edu.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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