Cloning and characterization of erythroid-specific DNase I-hypersensitive site in human rhesus-associated glycoprotein gene.
Rhesus-associated glycoprotein is a critical co-factor in the expression of rhesus blood group antigens. We identified and cloned an erythroid-specific major DNase I-hypersensitive site located about 10 kilobases upstream from the translation start site of the RHAG gene. A short core enhancer sequence of 195 base pairs that corresponded with the major hypersensitive site and possessed position- and orientation-independent enhancer activity in K562 cells. In vitro DNase I footprint analysis revealed four protected regions in the core enhancer; two GATA motifs, an Ets-like motif and an unknown motif. The GATA motifs bound GATA-1 and mutagenesis analysis revealed that the proximal one is critical for the enhancing activity. Homology plot analysis using the 5' sequence of the mouse RHAG gene revealed four homologous stretches and multiple insertions of repetitive sequences among them; four LINE/L1 and four Alu in the human and as well as one LINE/L1 and one LTR/MaLR in the mouse gene. The highly conservative enhancer region was flanked by SINE and LINE/L1 in both species. These results suggest that the 5'-flanking sequence of RHAG gene is a preferable target sequence for retroviral transposition and that the enhancer was inserted in the same manner, resulting in the acquisition of erythroid dominant expression.
Iwamoto
S
S
Department of Legal Medicine and Human Genetics, Jichi Medical School, Tochigi 329-0498, Japan. siwamoto@ms.jichi.ac.jp
Suganuma
H
H
Kamesaki
T
T
Omi
T
T
Okuda
H
H
Kajii
E
E
D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins; D003001:Cloning, Molecular; D018983:DNA Footprinting; D017931:DNA Primers; D003850:Deoxyribonuclease I / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D012204:Rh-Hr Blood-Group System / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D001798:Blood Proteins; D017931:DNA Primers; D008562:Membrane Glycoproteins; C498523:RHAG protein, human; D012204:Rh-Hr Blood-Group System; C401210:Rhag protein, mouse; D003850:Deoxyribonuclease I