Juxtaposition of the HPFH2 enhancer is not sufficient to reactivate the gamma-globin gene in adult erythropoiesis.
Previous studies have suggested that juxtaposition of a downstream enhancer to the fetal gamma-globin gene results in reactivation of the gamma-gene in adult erythrocytes of individuals with hereditary persistence of fetal hemoglobin (HPFH). To test the hypothesis in a much stricter basis, we produced beta locus YAC transgenic mice carrying an exact beta locus replicate of a deletional HPFH mutation, HPFH 2. Although the gamma-globin gene was expressed in the HPFH 2/beta locus YAC (HPFH2/YAC) transgenic mice in the early stage of development, it was completely silenced in the adult mice. The failure of gamma-gene reactivation by the juxtaposed HPFH2 enhancer contradicts the results of previous studies. We speculate that the discrepant results reflect differences in the distance between the locus of region (LCR) and the gamma-globin gene characteristic of the plasmid, cosmid or YAC constructs used for production of transgenic mice. The difference in the phenotype of the HPFH2/YAC transgenic mice and the humans with HPFH2 mutation suggests that in addition to juxtaposition of HPFH enhancers, the upstream region that is absent in the beta-YAC construct might be involved in gamma-gene reactivation in HPFH individuals. The DNase I hypersensitive sites of the LCR were well formed and the chromatin histones were acetylated. A moderate level of pol II binding was detected in the LCR, despite the fact that no transcription occurred in the globin-genes of the adult HPFH2/YAC transgenic mice. The results suggest that formation of the LCR chromatin structure in erythroid cells is independent of globin-gene transcription in the locus.
Xiang
Ping
P
Division of Medical Genetics, Department of Medicine, University of Washington, 98195, USA.
Han
Hemei
H
Barkess
Grainne
G
Olave
Ivan
I
Fang
Xiangdong
X
Yin
Wenxuan
W
Stamatoyannopoulos
George
G
Li
Qiliang
Q
D000107:Acetylation; D000818:Animals; D002843:Chromatin / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D004257:DNA Polymerase II / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D041905:Erythroid Cells / Q000378:metabolism; D004920:Erythropoiesis / Q000235:genetics*; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D005818:Genetic Engineering; D005914:Globins / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D006801:Humans; D019901:Locus Control Region / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D019076:Transgenes / Q000235:genetics
D002843:Chromatin; D006657:Histones; D005914:Globins; D004257:DNA Polymerase II