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pubmed-article:11973310pubmed:abstractTextRegulatory DNA from the Drosophila gene engrailed causes silencing of a linked reporter gene (mini-white) in transgenic Drosophila. This silencing is strengthened in flies homozygous for the transgene and has been called "pairing-sensitive silencing." The pairing-sensitive silencing activities of a large fragment (2.6 kb) and a small subfragment (181 bp) were explored. Since pairing-sensitive silencing is often associated with Polycomb group response elements (PREs), we tested the activities of each of these engrailed fragments in a construct designed to detect PRE activity in embryos. Both fragments were found to behave as PREs in a bxd-Ubx-lacZ reporter construct, while the larger fragment showed additional silencing capabilities. Using the mini-white reporter gene, a 139-bp minimal pairing-sensitive element (PSE) was defined. DNA mobility-shift assays using Drosophila nuclear extracts suggested that there are eight protein-binding sites within this 139-bp element. Mutational analysis showed that at least five of these sites are important for pairing-sensitive silencing. One of the required sites is for the Polycomb group protein Pleiohomeotic and another is GAGAG, a sequence bound by the proteins GAGA factor and Pipsqueak. The identity of the other proteins is unknown. These data suggest a surprising degree of complexity in the DNA-binding proteins required for PSE function.lld:pubmed
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pubmed-article:11973310pubmed:authorpubmed-author:FujiokaMikiMlld:pubmed
pubmed-article:11973310pubmed:authorpubmed-author:JaynesJames...lld:pubmed
pubmed-article:11973310pubmed:authorpubmed-author:KassisJudith...lld:pubmed
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pubmed-article:11973310pubmed:pagination1561-71lld:pubmed
pubmed-article:11973310pubmed:dateRevised2011-5-4lld:pubmed
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pubmed-article:11973310pubmed:articleTitleA complex array of DNA-binding proteins required for pairing-sensitive silencing by a polycomb group response element from the Drosophila engrailed gene.lld:pubmed
pubmed-article:11973310pubmed:affiliationLaboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.lld:pubmed
pubmed-article:11973310pubmed:publicationTypeJournal Articlelld:pubmed
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