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Nucleic Acids Research Advance Access published online on November 20, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn926
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Gene regulation, chromation and genetics

Evidence for RNA synthesis in the intergenic region between enhancer and promoter and its inhibition by insulators in Drosophila melanogaster

Nickolai A. Tchurikov*, Olga V. Kretova, Evgenia D. Moiseeva and Dmitri V. Sosin

Department of Genome Organization, Engelhardt Institute of Molecular Biology Russian Academy of Sciences, Moscow, 119991, Russia

*To whom correspondence should be addressed. Tel: +7 499 1359753; Fax: +7 499 1351405; Email: tchurikov{at}eimb.ru

Received August 5, 2008. Revised November 1, 2008. Accepted November 3, 2008.

Uncovering the nature of communication between enhancers, promoters and insulators is important for understanding the fundamental mechanisms that ensure appropriate gene expression levels. Here we describe an approach employing transient expression of genetic luciferase reporter gene constructs with quantitative RT–PCR analysis of transcription between an enhancer and Hsp70 promoter. We tested genetic constructs containing gypsy and/or Fab7 insulators in different orientations, and an enhancer from copia LTR-retroelement [(enh)copia]. A single gypsy or Fab7 insulator inserted between the promoter and enhancer in any polarity reduced enhancer action. A pair of insulators flanking the gene in any orientation exhibited increased insulation activity. We detected promoter-independent synthesis of non-coding RNA in the intergenic region of the constructs, which was induced by the enhancer in both directions and repressed by a single insulator or a pair of insulators. These results highlight the involvement of RNA-tracking mechanisms in the communications between enhancers and promoters, which are inhibited by insulators.


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