Interleukin-3 and erythropoietin cooperate in the regulation of the expression of erythroid-specific transcription factors during erythroid differentiation.
OBJECTIVE
To characterize how interleukin-3 and erythropoietin regulate cell fate by modulating the expression of lineage-specific transcription factors.


METHODS
This study analyzed mRNA and protein levels, gene transcription rates, and mRNA and protein stabilities of erythroid-specific transcription factors in lineage-restricted cells derived from the 32D cell line cultured either in interleukin-3 or erythropoietin.


RESULTS
Erythroid 32D subclones expressed levels of Idl, Gata-2, and Scl comparable and levels of Eklf and Gata-1 higher than those expressed by myeloid subclones. While maintained in interleukin-3, erythroid cells remained immature despite their high expression of Gata-1, Gata-2, Scl, Eklf, and Idl. Switching the erythroid cells to erythropoietin induced cell maturation (within 48 hours) and reduced expression of Gata-2 and Idl (in 24 hours) but did not alter the expression of Gata-1. The effects of interleukin-3 were mostly mediated by increases in transcription rates (Scl and Gata-2), and that of erythropoietin was apparently due to increased mRNA and protein (Gata-1, Scl, and Eklf) stability. In particular, erythropoietin increased the stability of the processed and transcriptionally more active form of GATA-1 protein.


CONCLUSIONS
These results suggest that interleukin-3 and erythropoietin cooperate to establish the lineage-specific transcription factor milieu of erythroid cells: interleukin-3 regulates mainly gene transcription and erythropoietin consistently increases mRNA and protein stability.
Ghinassi
Barbara
B
Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome, Italy.
Verrucci
Maria
M
Jelicic
Katija
K
Di Noia
Antonella
A
Migliaccio
Giovanni
G
Migliaccio
Anna Rita
AR
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D041905:Erythroid Cells / Q000166:cytology / Q000187:drug effects*; D050981:Erythroid-Specific DNA-Binding Factors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology / Q000502:physiology*; D020869:Gene Expression Profiling; D051798:Inhibitor of Differentiation Protein 1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D007377:Interleukin-3 / Q000494:pharmacology / Q000502:physiology*; D051379:Mice; D010641:Phenotype; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013329:Structure-Activity Relationship; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D013997:Time Factors
D051792:Basic Helix-Loop-Helix Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D004921:Erythropoietin; D051798:Inhibitor of Differentiation Protein 1; D007377:Interleukin-3; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; C497455:Idb1 protein, mouse; C489366:Tal1 protein, mouse