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Replacement of a Drosophila Polycomb response element core, and in situ analysis of its DNA motifs

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Abstract

Long-term repression of homeotic genes in the fruit fly is accomplished by proteins of the Polycomb Group, acting at Polycomb response elements (PREs). Here we use gene conversion to mutate specific DNA motifs within a PRE to test their relevance, and we exchange PREs to test their specificity. Previously we showed that removal of a 185 bp core sequence from the bithoraxoid PRE of the bithorax complex results in posteriorly directed segmental transformations. Mutating multiple binding sites for either the PHO or the GAF proteins separately in the core bithoraxoid PRE resulted in only rare and subtle transformations in adult flies. However, when both sets of sites were mutated, the transformations were similar in strength and penetrance to those caused by the deletion of the 185 bp core region. In contrast, mutating the singly occurring binding site of another DNA-binding protein, DSP1 (reportedly essential for PRE-activity), had no similar effect in combination with mutated PHO or GAF sites. Two minimal PREs from other segment-specific regulatory domains of the bithorax complex could substitute for the bithoraxoid PRE core. Our in situ analysis suggests that core PREs are interchangeable, and the cooperation between PHO and GAF binding sites is indispensable for silencing.

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Acknowledgments

We thank Anna Rehák, Edina Ördög, Anikó Berente, Ildikó Krausz and Margit Pál for the technical help, József Mihály and Henrik Gyurkovics for discussions and critical reading of the manuscript. This work was supported by a grant from the National Institutes of Health (W.B.) and a Fogarty International Award (L.S. and W.B.). G.K. was supported by the Dr. Rollin D. Hotchkiss Foundation.

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Authors and Affiliations

  1. Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Szeged, 6701, Hungary

    Gabriella Kozma & László Sipos

  2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA

    Welcome Bender

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  1. Gabriella Kozma
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  2. Welcome Bender
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  3. László Sipos
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Correspondence to László Sipos.

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Communicated by S. Hohmann.

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Kozma, G., Bender, W. & Sipos, L. Replacement of a Drosophila Polycomb response element core, and in situ analysis of its DNA motifs. Mol Genet Genomics 279, 595–603 (2008). https://doi.org/10.1007/s00438-008-0336-3

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