title: Spacer promoters are orientation-dependent activators of pre-rRNA transcription in Drosophila melanogaster.
abstract: In Drosophila melanogaster, 240-base-pair (bp) repeats, clustered in tandem arrays within the ribosomal DNA nontranscribed spacer region, include sites of RNA polymerase I-dependent transcription initiation and elements that stimulate the rate of transcription from the downstream precursor rRNA (pre-rRNA) promoter. We have analyzed the in vivo transcriptional activity of a large set of recombinant constructs in which tandem arrays of distinct segments derived from a 240-bp repeat were inserted upstream of the pre-rRNA promoter. The results indicate that activating spacer elements are confined to a region of 70 bp. Enhancing units overlap with spacer promoters, since DNA segments that stimulate transcription at the gene promoter also efficiently drive transcription initiation. The finding that artificial spacer arrays invariably stimulate pre-rRNA transcription initiation in an orientation-dependent fashion suggest that spacer-initiated transcription is involved in the enhancement process. The minimal spacer activating segment includes a perfect copy of a core domain of the gene promoter extending from -24 to +10 flanked by poorly homologous upstream DNA sequences. Spacer and gene promoters are functionally interchangeable as activating units. However, the different combination of DNA elements within the two determines a functional hierarchy, as only the pre-rRNA promoter is responsive to the stimulatory action of upstream units.
authors.affiliation: International Institute of Genetics and Biophysics, Consiglio Nazionale delle Ricerche, Naples, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Grimaldi
authors.forename: P
authors.initials: P
authors.lastname: Fiorentini
authors.forename: P P
authors.initials: PP
authors.lastname: Di Nocera
mesh_terms: D000818:Animals; D001483:Base Sequence; D004275:DNA, Ribosomal / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012318:RNA Polymerase I / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics*; D012335:RNA, Ribosomal / Q000235:genetics; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D004275:DNA, Ribosomal; D012322:RNA Precursors; D012335:RNA, Ribosomal; D012318:RNA Polymerase I