MafK/NF-E2 p18 is required for beta-globin genes activation by mediating the proximity of LCR and active beta-globin genes in MEL cell line.
Evidences indicate that locus control region (LCR) of beta-globin spatially closes to the downstream active gene promoter to mediate the transcriptional activation by looping. DNA binding proteins may play an important role in the looping formation. NF-E2 is one of the key transcription factors in beta-globin gene transcriptional activation. To shed light on whether NF-E2 is involved in this process, DS19MafKsiRNA cell pools were established by specifically knocked down the expression of MafK/NF-E2 p18, one subunit of NF-E2 heterodimer. In the above cell pools, it was observed that the occupancy efficiency of NF-E2 on beta-globin gene locus and the expression level of beta-globin genes were decreased. H3 acetylation, H3-K4 methylation and the deposition of RNA polymerase II, but not the recruitment of GATA-1, were also found reduced at the beta-globin gene cluster. Chromosome Conformation Capture (3C) assay showed that the cross-linking frequency between the main NF-E2 binding site HS2 and downstream structural genes was reduced compared to the normal cell. This result demonstrated that MafK/NF-E2 p18 recruitment was involved in the physical proximity of LCR and active beta-globin genes upon beta-globin gene transcriptional activation.
Du
Mei-Jun
MJ
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, PR China.
Lv
Xiang
X
Hao
De-Long
DL
Zhao
Guo-Wei
GW
Wu
Xue-Song
XS
Wu
Feng
F
Liu
De-Pei
DP
Liang
Chih-Chuan
CC
D000818:Animals; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004257:DNA Polymerase II / Q000502:physiology; D050982:GATA1 Transcription Factor / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology*; D020868:Gene Silencing; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D019901:Locus Control Region / Q000502:physiology*; D050985:MafK Transcription Factor / Q000502:physiology*; D051379:Mice; D034622:RNA Interference
D050982:GATA1 Transcription Factor; C494689:Gata1 protein, mouse; D006657:Histones; D050985:MafK Transcription Factor; C494696:Mafk protein, mouse; D005914:Globins; D004257:DNA Polymerase II