High Fractional Occupancy of a Tandem Maf Recognition Element and Its Role in Long-Range β-Globin Gene Regulation.
Enhancers and promoters assemble protein complexes that ultimately regulate the recruitment and activity of RNA polymerases. Previous work has shown that at least some enhancers form stable protein complexes, leading to the formation of enhanceosomes. We analyzed protein-DNA interactions in the murine β-globin gene locus using the methyltransferase accessibility protocol for individual templates (MAPit). The data show that a tandem Maf recognition element (MARE) in locus control region (LCR) hypersensitive site 2 (HS2) reveals a remarkably high degree of occupancy during differentiation of mouse erythroleukemia cells. Most of the other transcription factor binding sites in LCR HS2 or in the adult β-globin gene promoter regions exhibit low fractional occupancy, suggesting highly dynamic protein-DNA interactions. Targeting of an artificial zinc finger DNA-binding domain (ZF-DBD) to the HS2 tandem MARE caused a reduction in the association of MARE-binding proteins and transcription complexes at LCR HS2 and the adult βmajor-globin gene promoter but did not affect expression of the βminor-globin gene. The data demonstrate that a stable MARE-associated footprint in LCR HS2 is important for the recruitment of transcription complexes to the adult βmajor-globin gene promoter during erythroid cell differentiation.
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Jared R
JR
Stees
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Mir A
MA
Hossain
Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
Tomoki
T
Sunose
Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
Yasushi
Y
Kudo
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Carolina E
CE
Pardo
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Nancy H
NH
Nabilsi
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Russell P
RP
Darst
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Rosha
R
Poudyal
Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan CREST, Japan Science and Technology Agency, Sendai, Miyagi, Japan.
Kazuhiko
K
Igarashi
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA sumingh@ufl.edu jbungert@ufl.edu.
Suming
S
Huang
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Michael P
MP
Kladde
Department of Biochemistry and Molecular Biology, Center for Epigenetics, Genetics Institute, UF Health Cancer Center, Powell-Gene Therapy Center, University of Florida, Gainesville, Florida, USA sumingh@ufl.edu jbungert@ufl.edu.
Jörg
J
Bungert
D000818:Animals; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000378:metabolism; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D015973:Gene Expression Regulation, Leukemic; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics* / Q000378:metabolism; D019901:Locus Control Region; D051379:Mice; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation; D016335:Zinc Fingers; D055544:beta-Globins / Q000235:genetics*
D014157:Transcription Factors; D055544:beta-Globins; D004247:DNA; D012319:RNA Polymerase II