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

• Published in: The Journal of biological chemistry 2001-08, Vol.276 (34), p.31786-31792
• Main Authors: McNamara-Schroeder, K J, Hennessey, R F, Harding, G A, Jensen, R C, Stumph, W E
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Culture Techniques; DNA Primers; DNA-Directed RNA Polymerases - genetics; DNA-Directed RNA Polymerases - metabolism; Drosophila - genetics; Drosophila melanogaster; Promoter Regions, Genetic; RNA, Small Nuclear - genetics; snRNA U1; snRNA U6; Substrate Specificity; Transfection; transgenic flies
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M101273200

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.