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dc.contributor.authorGiraldo, Patricia-
dc.contributor.authorMartínez, Antonio-
dc.contributor.authorRegales, Lucía-
dc.contributor.authorLavado, Alfonso J.-
dc.contributor.authorGarcía Cabezas, Miguel Ángel-
dc.contributor.authorAlonso, Ángel-
dc.contributor.authorBusturia, Ana-
dc.contributor.authorMontoliu, Lluís-
dc.identifier.citationNucleic Acids Res. 31(21): 6290-6305 (2003)en_US
dc.description16 pages, 6 figures.-- PMID: 14576318 [PubMed].-- PMCID: PMC275449.-- DDBJ/EMBL/GenBank accession nos X76647, AF364302.-- Supplementary Material available.en_US
dc.description.abstractLocus control regions (LCRs) are complex high-order chromatin structures harbouring several regulatory elements, including enhancers and boundaries. We have analysed the mouse tyrosinase LCR functions, in vitro, in cell lines and, in vivo, in transgenic mice and flies. The LCR-core (2.1 kb), located at -15 kb and carrying a previously described tissue-specific DNase I hypersensitive site, operates as a transcriptional enhancer that efficiently transactivates heterologous promoters in a cell-specific orientation-independent manner. Furthermore, we have investigated the boundary activity of these sequences in transgenic animals and cells. In mice, the LCR fragment (3.7 kb) rescued a weakly expressed reference construct that displays position effects. In Drosophila, the LCR fragment and its core insulated the expression of a white minigene reporter construct from chromosomal position effects. In cells, sequences located 5' from the LCR-core displayed putative boundary activities. We have obtained genomic sequences surrounding the LCR fragment and found a LINE1 repeated element at 5'. In B16 melanoma and L929 fibroblast mouse cells, this element was found heavily methylated, supporting the existence of putative boundary elements that could prevent the spreading of condensed chromatin from the LINE1 sequences into the LCR fragment, experimentally shown to be in an open chromatin structure.en_US
dc.description.sponsorshipThis work was supported by funds from Spanish Ministry of Science and Technology (SMST) Bio97-0628, Bio2000-1653, FEDER 2FD1997-2059, and Laboratorios Dr. Esteve S.A. to L.M., and funds from SMST PB97-1207, BMC2001-2178 and Comunidad Autónoma de Madrid (CAM) 08.9/0002/98 to A.B. P.G. and L.R. are recipients of PhD fellowships from SMST. A.L. and A.G.D. are recipients from PhD fellowships from CAM.en_US
dc.format.extent454726 bytes-
dc.publisherOxford University Pressen_US
dc.subjectTyrosinase (Tyr)en_US
dc.subjectLocus control region (LCR)en_US
dc.subjectBoundary activityen_US
dc.subjectGene Expressiónen_US
dc.titleFunctional dissection of the mouse tyrosinase locus control region identifies a new putative boundary activityen_US
dc.description.peerreviewedPeer revieweden_US
dc.contributor.funderMinisterio de Ciencia y Tecnología (España)-
dc.contributor.funderComunidad de Madrid-
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