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Nucleic Acids Research 2005 33(20):6579-6586; doi:10.1093/nar/gki972
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Published online 27 November 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Article

The PSEA promoter element of the Drosophila U1 snRNA gene is sufficient to bring DmSNAPc into contact with 20 base pairs of downstream DNA

Hsien-Tsung Lai, Hsiang Chen, Cheng Li1, Kathleen J. McNamara-Schroeder1 and William E. Stumph1,*

Department of Biology, San Diego State University San Diego, CA 92182-1030, USA 1Department of Chemistry and Biochemistry, San Diego State University San Diego, CA 92182-1030, USA

*To whom correspondence should be addressed. Tel: +1 619 594 5575; Fax: +1 619 594 4634; Email: wstumph{at}sciences.sdsu.edu

Received September 11, 2005. Revised November 3, 2005. Accepted November 3, 2005.

Most of the major spliceosomal small nuclear RNAs (snRNAs) (i.e. U1, U2, U4 and U5) are synthesized by RNA polymerase II (pol II). In Drosophila melanogaster, the 5'-flanking DNA of these genes contains two conserved elements: the proximal sequence element A (PSEA) and the proximal sequence element B (PSEB). The PSEA is essential for transcription and is recognized by DmSNAPc, a multi-subunit protein complex. Previous site-specific protein–DNA photo-cross-linking assays demonstrated that one of the subunits of DmSNAPc, DmSNAP43, remains in close contact with the DNA for 20 bp beyond the 3' end of the PSEA, a region that contains the PSEB. The current work demonstrates that mutation of the PSEB does not abolish the cross-linking of DmSNAP43 to the PSEB. Thus the U1 PSEA alone is capable of bringing DmSNAP43 into close contact with this downstream DNA. However, mutation of the PSEB perturbs the cross-linking pattern. In concordance with these findings, PSEB mutations result in a 2- to 4-fold reduction in U1 promoter activity when assayed by transient transfection.

Present addresses: Hsiang Chen, Illumina, Inc., 9885 Towne Centre Drive, San Diego CA 92121-1975, USA

Cheng Li, Department of Immunology, Room222, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA


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