The PSEA promoter element of the Drosophila U1 snRNA gene is sufficient to bring DmSNAPc into contact with 20 base pairs of downstream DNA
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...
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Published in | Nucleic acids research Vol. 33; no. 20; pp. 6579 - 6586 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
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England
Oxford University Press
01.01.2005
Oxford Publishing Limited (England) |
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Abstract | 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. |
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AbstractList | 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. 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. |
Author | Chen, Hsiang Lai, Hsien-Tsung Li, Cheng Stumph, William E. McNamara-Schroeder, Kathleen J. |
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Notes | To whom correspondence should be addressed. Tel: +1 619 594 5575; Fax: +1 619 594 4634; Email: wstumph@sciences.sdsu.edu local:gki972 istex:00C8F4EF839DD082266A7B85F7096FB551ABA568 ark:/67375/HXZ-TL1QZRMD-V ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 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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SubjectTerms | Animals Base Pairing Cells, Cultured DNA - chemistry DNA - metabolism DNA-Binding Proteins - metabolism Drosophila melanogaster - genetics Drosophila Proteins - metabolism Mutation Promoter Regions, Genetic Response Elements RNA, Small Nuclear - genetics TATA Box Transcription Factors - metabolism Transcriptional Activation |
Title | The PSEA promoter element of the Drosophila U1 snRNA gene is sufficient to bring DmSNAPc into contact with 20 base pairs of downstream DNA |
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