Multiple interactions between regulatory regions are required to stabilize an active chromatin hub.
The human beta-globin locus control region (LCR) is required for the maintenance of an open chromatin configuration of the locus. It interacts with the genes and the hypersensitive regions flanking the locus to form an active chromatin hub (ACH) transcribing the genes. Proper developmental control of globin genes is largely determined by gene proximal regulatory sequences. Here, we provide the first functional evidence of the role of the most active sites of the LCR and the promoter of the beta-globin gene in the maintenance of the ACH. When the human beta-globin gene promoter is deleted in the context of a full LCR, the ACH is maintained with the beta-globin gene remaining in proximity. Additional deletion of hypersensitive site HS3 or HS2 of the LCR shows that HS3, but not HS2, in combination with the beta-globin promoter is crucial for the maintenance of the ACH at the definitive stage. We conclude that multiple interactions between the LCR and the beta-globin gene are required to maintain the appropriate spatial configuration in vivo.
Patrinos
George P
GP
Erasmus University Medical Center, Faculty of Medicine and Health Sciences, MGC Department of Cell Biology and Genetics, Rotterdam, 3000 DR, The Netherlands.
de Krom
Mariken
M
de Boer
Ernie
E
Langeveld
An
A
Imam
A M Ali
AM
Strouboulis
John
J
de Laat
Wouter
W
Grosveld
Frank G
FG
D000818:Animals; D002843:Chromatin / Q000648:ultrastructure*; D005914:Globins / Q000235:genetics*; D006801:Humans; D019901:Locus Control Region / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017384:Sequence Deletion; D014158:Transcription, Genetic
D002843:Chromatin; D005914:Globins