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Genetics, Vol. 168, 1371-1384, November 2004, Copyright © 2004
doi:10.1534/genetics.104.029561
The Enhancer-Blocking Activity of the Fab-7 Boundary From the Drosophila Bithorax Complex Requires GAGA-Factor-Binding Sites
Susan Schweinsberg*,
Kirsten Hagstrom*,
,
Daryl Gohl*,
Paul Schedl*,1,
Ram P. Kumar
,
Rakesh Mishra and
Francois Karch
* Department of Molecular Biology, Princeton University, Princeton, New Jersey 08540
Centre for Cellular and Molecular Biology, Hyderabad 500 007, India
Département de Zoologie et Biologie Animale, Université de Genève, 1211 Genève 4, Switzerland
Program in Molecular Medicine and Program in Cell Dynamics, University of Massachusetts Medical School, Worcester, Massachusetts 01605
1 Corresponding author: Department of Molecular Biology, Lewis Thomas Labs, Washington Rd., Princeton University, Princeton, NJ 08544.
E-mail: pschedl{at}molbio.princeton.edu
In the work reported here we have analyzed the role of the GAGA factor [encoded by the Trithorax-like (Trl) gene] in the enhancer-blocking activity of Frontabdominal-7 (Fab-7), a domain boundary element from the Drosophila melanogaster bithorax complex (BX-C). One of the three nuclease hypersensitive sites in the Fab-7 boundary, HS1, contains multiple consensus-binding sequences for the GAGA factor, a protein known to be involved in the formation and/or maintenance of nucleosome-free regions of chromatin. GAGA protein has been shown to localize to the Fab-7 boundary in vivo, and we show that it recognizes sequences from HS1 in vitro. Using two different transgene assays we demonstrate that GAGA-factor-binding sites are necessary but not sufficient for full Fab-7 enhancer-blocking activity. We show that distinct GAGA sites are required for different enhancer-blocking activities at different stages of development. We also show that the enhancer-blocking activity of the endogenous Fab-7 boundary is sensitive to mutations in the gene encoding the GAGA factor Trithorax-like.
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