Cell-cycle-dependent regulation of erythropoietin receptor gene

Blood. 1997 Feb 15;89(4):1182-8.

Abstract

To understand the regulatory mechanism of erythropoietin (EPO) receptor (EPOR) gene expression, the effect of EPO on the steady-state level of EPOR mRNA was examined using the human EPO-dependent cell line UT-7 as a model system. We found that the treatment of UT-7 cells with EPO resulted in a transient decrease of the EPOR mRNA level. This transient downregulation was also induced by stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF), another stimulator of UT-7 cell growth. These results raised the possibility that EPOR gene expression is in part related to cell growth. Moreover, it was found that EPO-induced downregulation of EPOR mRNA level was preceded by a transient downregulation of GATA-1 mRNA. To examine the relationship between the expression of EPOR, GATA-1, and GATA-2 mRNA levels and the cell cycle, logarithmically growing UT-7 cells were centrifugically fractionated according to the cell-cycle phase. Both EPOR and GATA-1 mRNA levels, but not the GATA-2 mRNA level, concomitantly decreased at the G0/G1 phase and increased at the S and G2/M phases. An electrophoretic mobility shift assay (EMSA) showed that in EPO-stimulated UT-7 cells, the dynamic changes in EPOR gene expression paralleled the GATA-1 DNA-binding activity to the oligonucleotide probe containing a GATA-binding site located at the promoter region of the EPOR gene. These findings suggest that the regulation of EPOR mRNA level is mainly associated with GATA-1 gene expression in UT-7 cells undergoing proliferation, and that these serial events are under the control of, or related to, the cell cycle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle*
  • Cell Division
  • Cell Line
  • Cell Separation
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics
  • Erythroid-Specific DNA-Binding Factors
  • Erythropoietin / pharmacology
  • GATA1 Transcription Factor
  • GATA2 Transcription Factor
  • Gene Expression Regulation, Leukemic* / drug effects
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Leukemia, Megakaryoblastic, Acute / pathology
  • Models, Genetic
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Receptors, Erythropoietin / biosynthesis
  • Receptors, Erythropoietin / genetics*
  • Recombinant Proteins / pharmacology
  • Resting Phase, Cell Cycle
  • Transcription Factors / biosynthesis
  • Transcription Factors / genetics

Substances

  • DNA-Binding Proteins
  • Erythroid-Specific DNA-Binding Factors
  • GATA1 Transcription Factor
  • GATA1 protein, human
  • GATA2 Transcription Factor
  • GATA2 protein, human
  • Neoplasm Proteins
  • RNA, Messenger
  • Receptors, Erythropoietin
  • Recombinant Proteins
  • Transcription Factors
  • Erythropoietin
  • Granulocyte-Macrophage Colony-Stimulating Factor