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Context Differences Reveal Insulator and Activator Functions of a Su(Hw) Binding Region
Journal article   Open access   Peer reviewed

Context Differences Reveal Insulator and Activator Functions of a Su(Hw) Binding Region

Alexey A Soshnev, Xingguo Li, Misty D Wehling and Pamela K Geyer
PLoS genetics, Vol.4(8), pp.e1000159-e1000159
08/2008
DOI: 10.1371/journal.pgen.1000159
PMCID: PMC2493044
PMID: 18704163
url
https://doi.org/10.1371/journal.pgen.1000159View
Published (Version of record) Open Access

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 - a chaete ( ac ) intergenic R NA. 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. Insulators are conserved genomic elements that define domains of independent transcription. One class of insulators in the Drosophila genome are defined by the binding of the Su(Hw) protein, with the gypsy insulator representing the classic Su(Hw)-dependent insulator. Su(Hw) associates with hundreds of non- gypsy regions distributed throughout the genome that differ in sequence and organization from the gypsy insulator. To gain insights into the role of Su(Hw) in genome organization, we defined the properties of the first non- gypsy Su(Hw) binding region identified, 1A-2. Our studies reveal differences in 1A-2 activity, depending on the context tested. We show that 1A-2 is an insulator in enhancer blocking studies but functions as a transcriptional activator within the natural genomic location. Our findings are reminiscent of properties of binding regions that associate with the vertebrate CTCF protein, which have defined insulator, activator, and repressor functions. Finally, our studies indicate that a noncoding RNA gene may contribute to independent transcriptional regulation in the genome.
Gene Function Transcription Initiation and Activation Gene Expression Molecular Biology Chromosome Structure Genetics and Genomics Functional Genomics Biochemistry Nuclear Structure and Function Transcription and Translation

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