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Molecular and Cellular Biology, July 1999, p. 5189-5202, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Trithorax- and Polycomb-Group Response Elements within an Ultrabithorax Transcription Maintenance Unit Consist of Closely Situated but Separable Sequencesdagger

Sergei Tillib, Svetlana Petruk, Yurii Sedkov, Alexander Kuzin, Miki Fujioka, Tadaatsu Goto, and Alexander Mazo*

Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107

Received 15 December 1998/Returned for modification 21 January 1999/Accepted 13 April 1999

In Drosophila, two classes of genes, the trithorax group and the Polycomb group, are required in concert to maintain gene expression by regulating chromatin structure. We have identified Trithorax protein (TRX) binding elements within the bithorax complex and have found that within the bxd/pbx regulatory region these elements are functionally relevant for normal expression patterns in embryos and confer TRX binding in vivo. TRX was localized to three closely situated sites within a 3-kb chromatin maintenance unit with a modular structure. Results of an in vivo analysis showed that these DNA fragments (each ~400 bp) contain both TRX- and Polycomb-group response elements (TREs and PREs) and that in the context of the endogenous Ultrabithorax gene, all of these elements are essential for proper maintenance of expression in embryos. Dissection of one of these maintenance modules showed that TRX- and Polycomb-group responsiveness is conferred by neighboring but separable DNA sequences, suggesting that independent protein complexes are formed at their respective response elements. Furthermore, we have found that the activity of this TRE requires a sequence (~90 bp) which maps to within several tens of base pairs from the closest neighboring PRE and that the PRE activity in one of the elements may require a binding site for PHO, the protein product of the Polycomb-group gene pleiohomeotic. Our results show that long-range maintenance of Ultrabithorax expression requires a complex element composed of cooperating modules, each capable of interacting with both positive and negative chromatin regulators.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Room 485, Jefferson Alumni Hall, 1020 Locust St., Philadelphia, PA 19107. Phone: (215) 503-4785. Fax: (215) 923-7144. E-mail: mazo{at}lac.jci.tju.edu.

dagger This work is dedicated to the memory of Tadaatsu Goto.

Molecular and Cellular Biology, July 1999, p. 5189-5202, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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