Globin Messenger RNA Activity in Erythroid Precursor Cells and the Effect of Erythropoietin

  1. Masaaki Terada,
  2. Linda Cantor,
  3. Salvatore Metafora,
  4. Richard A. Rifkind,
  5. Arthur Bank, and
  6. Paul A. Marks
  1. Department of Medicine, Columbia University, College of Physicians & Surgeons, New York, N.Y. 10032
  2. Department of Human Genetics and Development, Columbia University, College of Physicians & Surgeons, New York, N.Y. 10032

Abstract

Cell populations enriched for erythroid precursor cells were fractionated from 13-day fetal-mouse livers by a method of immune hemolysis. These preparations of precursors, contaminated by less than 7% hemoglobinized erythroblasts, synthesize globin at a rate less than 6% that of the unfractionated liver erythroid cell population. RNA was isolated from these precursor cells and assayed for globin mRNA activity in a cell-free system from Krebs ascites tumor. The 6-16S RNA fraction from precursor cells has less than 5% of the globin mRNA activity of RNA isolated from unfractionated populations. Precursor cells incubated with erythropoietin show an increment in the rate of synthesis of globin only after 5-10 hr of incubation. After 10 hr of culture with this hormone, precursor cells show a 6- to 10-fold increase in globin mRNA activity. These results suggest that the precursor cells of hepatic erythropoiesis, responsive to erythropoietin, do not contain globin mRNA in a biologically active form. Erythropoietin-induced differentiation of these cells to erythroblasts is associated with an increase in globin mRNA.

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