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Identification of the proximal erythroid promoter region of the mouse anion
exchanger gene
KE Sahr, BP Daniels and M Hanspal
Department of Biomedical Research, St Elizabeth's Medical Center of Boston,
Tufts University School of Medicine, MA 02135, USA.
The AE1 gene is expressed in erythrocytes and the A-type intercalated cells
of the kidney distal collecting duct. Although the 5' end of the principal
transcript expressed in murine erythroid cells has previously been mapped
to a cluster of transcription start sites located immediately upstream of
exon 1, the 5' end of the mouse kidney transcript has not been identified.
Using the anchored polymerase chain reaction technique to analyze mouse
kidney AE1 mRNA, we identified an internal transcription start site located
within erythroid intron 3. This site defines an exon of 37 nucleotides that
forms the 5' end of the mouse kidney AE1 transcript. AE1 transcripts
beginning at this internal start site could not be detected in RNA isolated
from purified erythroid progenitor cells or from erythroid cells undergoing
erythropoietin-dependent terminal maturation, although transcripts derived
from the upstream site were abundant, indicating that only the upstream
promoter is active during erythropoiesis. Transient expression of reporter
constructs in erythroid and nonerythroid cell lines identified a proximal
upstream region of approximately 135 nucleotides that was active as a basal
promoter. However, an additional approximately 200 nucleotides of upstream
sequence was required for induced levels of activity in erythroid cells.
Although our functional approach does not yet indicate the precise
sequences required for erythroid induction, the AE1 gene upstream region
contains potential GATA sites at -154, -141, and -60; an E-box at -163;
CACCC or GGTGG motifs at -188, -121, and -88; and an AP-1/NF-E2-like site
at -42.
Volume 88,
Issue 12,
pp. 4500-4509,
12/15/1996
Copyright © 1996 by The American Society of Hematology
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