The murine beta-globin locus control region regulates the rate of transcription but not the hyperacetylation of histones at the active genes.
Locus control regions (LCRs) are defined by their ability to confer high-level tissue-specific expression to linked genes in transgenic assays. Previously, we reported that, at its native site, the murine beta-globin LCR is required for high-level beta-globin gene expression, but is not required to initiate an open chromatin conformation of the locus. To further investigate the mechanism of LCR-mediated transcriptional enhancement, we have analyzed allele-specific beta-globin expression and the pattern of histone acetylation in the presence and absence of the LCR. In single cells from mice heterozygous for a deletion of the LCR, beta-globin expression from the LCR-deleted allele is consistently low ( approximately 1-4% of wild type). Thus, the endogenous LCR enhances globin gene expression by increasing the rate of transcription from each linked allele rather than by increasing the probability of establishing transcription per se. Furthermore, in erythroid cells from mice homozygous for the highly expressing wild-type beta-globin locus, hyperacetylation of histones H3 and H4 is localized to the LCR and active genes. In mice homozygous for the LCR deletion reduced histone hyperacetylation is observed in LCR proximal sequences; however, deletion of the LCR has no effect on the localized hyperacetylation of the genes. Together, our results suggest that, in its native genomic context, the LCR follows the rate model of enhancer function, and that the developmentally specific hyperacetylation of the globin genes is independent of both the rate of transcription and the presence of the LCR.
Schübeler
D
D
Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Groudine
M
M
Bender
M A
MA
D000107:Acetylation; D000483:Alleles; D000818:Animals; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics*; D006657:Histones / Q000235:genetics*; D019901:Locus Control Region; D051379:Mice; D016133:Polymerase Chain Reaction; D011336:Probability; D014158:Transcription, Genetic
D006657:Histones; D005914:Globins