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Cloning of the gene encoding the human erythropoietin receptor
L Maouche, C Tournamille, C Hattab, G Boffa, JP Cartron and S Chretien
Institut National de Transfusion Sanguine, Paris, France.
The genomic and complementary DNAs of the human erythropoietin receptor
(hEpo-R) have been isolated and characterized from a genomic placental
library and from two cDNA libraries prepared from bone marrow and fetal
liver. The five different partial cDNAs isolated were aberrant in the
predicted reading frames as compared with the Epo-R protein sequence,
because all retained insert sequences that may represent splicing
intermediates (three clones), cloning artifact (one clone), or a new
sequence at a splice junction (one clone) of the gene. The cDNAs were used
to isolate several genomic clones encompassing the complete hEpo-R gene.
This gene, which encodes a 508-amino acid polypeptide chain of predicted
M(r) 55,000, is organized into eight exons spread over 6 kb of DNA and
exhibited a high degree of sequence homology (81.6% in the coding region)
and structural organization with its murine counterpart. Primer extension
analysis indicated that the transcription initiation site is located 141 bp
upstream of the initiation codon. Sequence homology 320 bp upstream of the
cap site was significantly lower (60%) and diverged completely further
upstream as compared with the murine gene. Similarly, the human and murine
sequences were largely divergent downstream of the stop codon, indicating
that a strong conservation during evolution was restricted to the coding
sequence of the Epo-R protein. The 320-bp region upstream of the cap site
does not contain the typical TATA or CAAT boxes present in many
tissue-specific genes, but does include potential binding sites for the
ubiquitous Sp1 and the erythroid-specific GATA-1 trans-activating factors.
These boxes are well conserved in sequence and position relative to the cap
site within the promoter region of the human and murine genes, but the
CACCC boxes present in the murine gene are absent in the human gene.
Volume 78,
Issue 10,
pp. 2557-2563,
11/15/1991
Copyright © 1991 by The American Society of Hematology
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