GATA-1 and Oct-1 are required for expression of the human α-hemoglobin-stabilizing protein gene

被引:33
作者
Gallagher, PG
Liem, RI
Wong, E
Weiss, MJ
Bodine, DM
机构
[1] Yale Univ, Dept Pediat, Sch Med, New Haven, CT 06520 USA
[2] Childrens Hosp Philadelphia, Philadelphia, PA 19104 USA
[3] Univ Penn, Philadelphia, PA 19104 USA
[4] NHGRI, Hematopoiesis Sect, NIH, Bethesda, MD 20892 USA
关键词
D O I
10.1074/jbc.M506062200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
alpha-Hemoglobin-stabilizing protein (AHSP) is an erythroid protein that binds and stabilizes alpha-hemoglobin during normal erythropoiesis and in pathological states of alpha-hemoglobin excess. AHSP has been proposed as a candidate gene in some Heinz body hemolytic anemias and as a modifier gene in the beta-thalassemia syndromes. To gain additional insight into the molecular mechanisms controlling the erythroid-specific expression of the AHSP gene and provide the necessary tools for further genetic studies of these disorders, we have initiated identification and characterization of the regulatory elements controlling the human AHSP gene. We mapped the 5'-end of the AHSP erythroid cDNA and cloned the 5'-flanking genomic DNA containing the putative AHSP gene promoter. In vitro studies using transfection of promoter/reporter plasmids in human tissue culture cell lines, DNase I footprinting analyses and gel mobility shift assays, identified an AHSP gene erythroid promoter with functionally important binding sites for GATA-1- and Oct-1-related proteins. In transgenic mice, a reporter gene directed by a minimal human AHSP promoter was expressed in bone marrow, spleen, and reticulocytes, but not in nonerythroid tissues. In vivo studies using chromatin immunoprecipitation assays demonstrated hyperacetylation of the promoter region and occupancy by GATA-1. The AHSP promoter is an excellent candidate region for mutations associated with decreased AHSP gene expression.
引用
收藏
页码:39016 / 39023
页数:8
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