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Prepublished online as a Blood First Edition Paper on October 31, 2002; DOI 10.1182/blood-2002-04-1217.
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Blood, 15 March 2003, Vol. 101, No. 6, pp. 2235-2242
HEMATOPOIESIS
The function of the bcl-x promoter in erythroid
progenitor cells
Cuixia Tian,
Paul Gregoli, and
Maurice Bondurant
From the Veterans Affairs and Vanderbilt
University Medical Centers, Nashville, TN.
The protein Bcl-xL is essential for survival of
erythroid progenitor cells, and it increases substantially during late
erythrocyte differentiation due to an increase of mRNA. We mapped the
transcription start sites of bcl-x mRNA in mouse and human
erythroblasts, and we analyzed the function of the mouse
bcl-x promoter by transient and stable transfection assays
in a mouse erythroid cell line using plasmids containing the
bcl-x promoter fused to a luciferase reporter gene. In
mouse erythroblasts, a cluster of start sites at positions  664,
655, and 644 relative to the ATG initiation codon account for
almost all transcripts. Human erythroblasts exhibit a start site at
654 that is homologous to the triplet in the mouse. A short sequence
element in the mouse bcl-x promoter that includes
nucleotides 1804 through 1734 was identified as very important for
transcription. This element also showed strong enhancerlike activity in
concert with the SV40 promoter in an enhancer test vector. Analyses of
mutations indicated that 2 short sequences within the element, about 15 base pair apart, are necessary for full enhancer activity. Gel shift
experiments with oligonucleotides representing these sequences revealed
specific binding of nuclear proteins from erythroblasts. Some of these
proteins are regulated during the late erythroid differentiation.
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