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• W2084659836 endingPage "2640" @default.
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• W2084659836 abstract "The human beta-globin genes are regulated by the locus control region (LCR), an element composed of multiple DNase I-hypersensitive sites (HS sites) located 5' to the genes. Various functional studies indicate that the LCR confers high-level, position-independent, and copy number-dependent expression to linked globin genes in transgenic mice. However, the structural basis for LCR function is unknown. Here we show that LCR HS sites can be reconstituted in an erythroid cell-specific manner on chromatin-assembled LCR templates in vitro. Surprisingly, HS2 and HS3 are also formed with erythroid proteins in the absence of chromatin assembly, indicating that sensitivity to nucleases is not simply a consequence of nucleosome reorganization. The generation of LCR HS sites in the absence of chromatin assembly leads to the formation of S1- and KMnO(4)-sensitive regions in HS2 and HS3. These sites are also sensitive to S1 nuclease in erythroid cells in vivo, suggesting a distorted DNA structure in the LCR core enhancer elements. Finally, we show that RNA polymerase II initiates transcription in the HS2 and HS3 core enhancer regions in vitro. Transcription in both HS2 and HS3 proceeds in a unidirectional manner. Taken together, the data suggest that erythroid proteins interact with the core enhancer elements, distort the DNA structure, and recruit polymerase II transcription complexes. These results further our understanding of the structural basis for LCR function and provide an explanation for why the LCR core regions are so extremely sensitive to nucleases in erythroid cells." @default.
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• W2084659836 date "2001-04-01" @default.
• W2084659836 modified "2023-09-23" @default.
• W2084659836 title "Reconstitution of Human β-Globin Locus Control Region Hypersensitive Sites in the Absence of Chromatin Assembly" @default.
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• W2084659836 doi "https://doi.org/10.1128/mcb.21.8.2629-2640.2001" @default.
• W2084659836 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/86894" @default.
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