Recruitment of transcription complexes to the beta-globin locus control region and transcription of hypersensitive site 3 prior to erythroid differentiation of murine embryonic stem cells.
Eukaryotic chromosomal DNA is densely packaged in the nucleus and organized into discrete domains of active and inactive chromatin. Gene loci that are activated during the process of cell differentiation undergo changes that result in modifications of specific histone tail residues and in loosening of chromatin structure. The beta-globin genes are expressed exclusively in erythroid cells. High-level expression of these genes is mediated by a locus control region (LCR), a powerful DNA regulatory element composed of several DNase I hypersensitive (HS) sites and located far upstream of the beta-globin genes. Here we show that RNA polymerase II and specific histone modifications that mark transcriptionally active chromatin domains are associated with the LCR core elements HS2 and HS3 in murine embryonic stem cells prior to differentiation along the erythroid lineage. At this stage HS3 is abundantly transcribed. After in vitro differentiation, RNA Polymerase II can also be detected at the embryonic epsilon- and adult beta-globin genes. These results are consistent with the hypothesis that activation of the beta-globin gene locus is initiated by protein complexes recruited to the LCR.
Levings
Padraic P
PP
Department of Biochemistry and Molecular Biology, University of Florida, Center for Mammalian Genetics, Shands Cancer Center, Powell Gene Therapy Center, Gainesville, Florida 32610, USA.
Zhou
Zhuo
Z
Vieira
Karen F
KF
Crusselle-Davis
Valerie J
VJ
Bungert
Jörg
J
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D019901:Locus Control Region; D046911:Macromolecular Substances; D051379:Mice; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014158:Transcription, Genetic
D002843:Chromatin; D006657:Histones; D046911:Macromolecular Substances; D005914:Globins; D012319:RNA Polymerase II