title: Context differences reveal insulator and activator functions of a Su(Hw) binding region.
abstract: Insulators are DNA elements that divide chromosomes into independent transcriptional domains. The Drosophila genome contains hundreds of binding sites for the Suppressor of Hairy-wing [Su(Hw)] insulator protein, corresponding to locations of the retroviral gypsy insulator and non-gypsy binding regions (BRs). The first non-gypsy BR identified, 1A-2, resides in cytological region 1A. Using a quantitative transgene system, we show that 1A-2 is a composite insulator containing enhancer blocking and facilitator elements. We discovered that 1A-2 separates the yellow (y) gene from a previously unannotated, non-coding RNA gene, named yar for y-achaete (ac) intergenic RNA. The role of 1A-2 was elucidated using homologous recombination to excise these sequences from the natural location, representing the first deletion of any Su(Hw) BR in the genome. Loss of 1A-2 reduced yar RNA accumulation, without affecting mRNA levels from the neighboring y and ac genes. These data indicate that within the 1A region, 1A-2 acts an activator of yar transcription. Taken together, these studies reveal that the properties of 1A-2 are context-dependent, as this element has both insulator and enhancer activities. These findings imply that the function of non-gypsy Su(Hw) BRs depends on the genomic environment, predicting that Su(Hw) BRs represent a diverse collection of genomic regulatory elements.
authors.affiliation: Program in Molecular and Cellular Biology, University of Iowa, Iowa City, Iowa, United States of America.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Soshnev
authors.forename: Xingguo
authors.initials: X
authors.lastname: Li
authors.forename: Misty D
authors.initials: MD
authors.lastname: Wehling
authors.forename: Pamela K
authors.initials: PK
authors.lastname: Geyer
mesh_terms: D000818:Animals; D001665:Binding Sites; D021901:DNA, Intergenic / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D049750:Genome, Insect; D038101:Insulator Elements; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D022661:RNA, Untranslated; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C423782:e(y)2 protein, Drosophila; C083738:su(Hw) protein, Drosophila