Cooperativeness of the higher chromatin structure of the beta-globin locus revealed by the deletion mutations of DNase I hypersensitive site 3 of the LCR.
High-level transcription of the globin genes requires the enhancement by a distant element, the locus control region (LCR). Such long-range regulation in vivo involves spatial interaction between transcriptional elements, with intervening chromatin looping out. It has been proposed that the clustering of the HS sites of the LCR, the active globin genes, as well as the remote 5' hypersensitive sites (HSs) (HS-60/-62 in mouse, HS-110 in human) and 3'HS1 forms a specific spatial chromatin structure, termed active chromatin hub (ACH). Here we report the effects of the HS3 deletions of the LCR on the spatial chromatin structure of the beta-globin locus as revealed by the chromatin conformation capture (3C) technology. The small HS3 core deletion (0.23 kb), but not the large HS3 deletion (2.3 kb), disrupted the spatial interactions among all the HS sites of the LCR, the beta-globin gene and 3'HS1. We have previously demonstrated that the large HS3 deletion barely impairs the structure of the LCR holocomplex, while the structure is significantly disrupted by the HS3 core deletion. Taken together, these results suggest that the formation of the ACH is dependent on a largely intact LCR structure. We propose that the ACH indeed is an extension of the LCR holocomplex.
Fang
Xiangdong
X
Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Xiang
Ping
P
Yin
Wenxuan
W
Stamatoyannopoulos
George
G
Li
Qiliang
Q
D000818:Animals; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D003850:Deoxyribonuclease I / Q000378:metabolism; D004900:Erythroblasts / Q000378:metabolism; D005914:Globins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019901:Locus Control Region; D051379:Mice; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction / Q000379:methods; D017384:Sequence Deletion; D013154:Spleen / Q000166:cytology; D014158:Transcription, Genetic
D002843:Chromatin; D005914:Globins; D003850:Deoxyribonuclease I