The iab-7 Polycomb Response Element Maps to a Nucleosome-Free Region of Chromatin and Requires Both GAGA and Pleiohomeotic for Silencing Activity

doi:10.1128/MCB.21.4.1311-1318.2001

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Molecular and Cellular Biology, February 2001, p. 1311-1318, Vol. 21, No. 4
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.4.1311-1318.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The iab-7 Polycomb Response Element Maps to a Nucleosome-Free Region of Chromatin and Requires Both GAGA and Pleiohomeotic for Silencing Activity

Rakesh K. Mishra,¹ Jozsef Mihaly,² Stéphane Barges,¹ Annick Spierer,¹ François Karch,¹ Kirsten Hagstrom,³ Susan E. Schweinsberg,³ and Paul Schedl³^,^*

Département de Zoologie et Biologie Animale, Université de Genève, 1211 Geneva 4, Switzerland¹; Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, 6701 Szeged, Hungary²; and Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544³

Received 3 August 2000/Returned for modification 25 September 2000/Accepted 17 October 2000

In the work reported here we have undertaken a functional dissection of a Polycomb response element (PRE) from the iab-7 cis-regulatorydomain of the Drosophila melanogaster bithorax complex (BX-C).Previous studies mapped the iab-7 PRE to an 860-bp fragment locatedjust distal to the Fab-7 boundary. Located within this fragmentis an ~230-bp chromatin-specific nuclease-hypersensitive regioncalled HS3. We have shown that HS3 is capable of functioning asa Polycomb-dependent silencer in vivo, inducing pairing-dependentsilencing of a mini-white reporter. The HS3 sequence containsconsensus binding sites for the GAGA factor, a protein implicatedin the formation of nucleosome-free regions of chromatin, andPleiohomeotic (Pho), a Polycomb group protein that is relatedto the mammalian transcription factor YY1. We show that GAGA andPho interact with these sequences in vitro and that the consensusbinding sites for the two proteins are critical for the silencingactivity of the iab-7 PRE invivo.

^* Corresponding author. Mailing address: Department of Molecular Biology, Princeton University, Princeton, NJ 08544. Phone:(609) 258-4979. Fax: (609) 258-1028. E-mail: pschedl{at}molbio.princeton.edu.

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