Functional and binding studies of HS3.2 of the beta-globin locus control region.
The distal locus control region (LCR) is required for high-level expression of the complex of genes (HBBC) encoding the beta-like globins of mammals in erythroid cells. Several major DNase hypersensitive sites (HSs 1-5) mark the LCR. Sequence conservation and direct experimental evidence have implicated sequences within and between the HS cores in function of the LCR. In this report we confirm the mapping of a minor HS between HS3 and HS4, called HS3.2, and show that sequences including it increase the number of random integration sites at which a drug resistance gene is expressed. We also show that nuclear proteins including GATA1 and Oct1 bind specifically to sequences within HS3.2. However, the protein Pbx1, whose binding site is the best match to one highly conserved sequence, does not bind strongly. GATA1 and Oct1 also bind in the HS cores of the LCR and to promoters in HBBC. Their binding to this minor HS suggests that they may be used in assembly of a large complex containing multiple regulatory sequences.
Molete
Joseph M
JM
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 206 Althouse Laboratory, University Park, PA 16802, USA
Petrykowska
Hanna
H
Sigg
Martin
M
Miller
Webb
W
Hardison
Ross
R
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D005914:Globins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D020014:K562 Cells; D019901:Locus Control Region / Q000235:genetics*; D008969:Molecular Sequence Data; D027702:Organic Cation Transporter 1 / Q000378:metabolism; D011485:Protein Binding; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; D027702:Organic Cation Transporter 1; D014157:Transcription Factors; D005914:Globins; D004247:DNA; D003851:Deoxyribonucleases