Abstract
Proper expression of the c-myb proto-oncogene is essential for definitive, but not primitive erythropoiesis. To examine the role of c-myb during adult erythropoiesis, we incubated purified murine colony-forming units (CFU-E) with a c-myb-specific antisense oligodeoxynucleotide (AS- oligo) in order to diminish expression levels. By western blot analysis, c- myb expression was reduced during the first seven hours of AS-oligo treatment as compared to untreated cells. We then quantitated the amount of heme synthesized in CFU-E treated with c-myb AS-oligo, a random sequence oligo or no oligo. No significant differences were seen in tile amount of heme synthesized during 42 hours of erythroid culture with either high levels (1 U/mL) or physiological levels (20 mU/mL) of Epo. In contrast, CFU-E treated with an AS-oligo directed toward mRNA encoding the first enzyme of the heme biosynthetic pathway in erythroid cells (d-aminolevulinate synthase) demonstrated a 65% reduction in the amount of heme synthesized. We conclude that the major role of c-myb during hematopoiesis must be in progenitor cells antecedent to the CFU-E stage and may possibly involve the establishment of the genetic program directing the formation of red blood cells.
Original language | English (US) |
---|---|
Pages (from-to) | 68-77 |
Number of pages | 10 |
Journal | Blood Cells, Molecules, and Diseases |
Volume | 25 |
Issue number | 1 |
DOIs | |
State | Published - Feb 1999 |
Keywords
- Antisense oligodeoxynucleotides
- C-myb
- CFU-E
- Erythropoietin
- Heme
ASJC Scopus subject areas
- Molecular Medicine
- Molecular Biology
- Hematology
- Cell Biology