Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1972 Dec;69(12):3575–3579. doi: 10.1073/pnas.69.12.3575

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

Masaaki Terada 1,2, Linda Cantor 1,2, Salvatore Metafora 1,2, Richard A Rifkind 1,2, Arthur Bank 1,2, Paul A Marks 1,2
PMCID: PMC389824  PMID: 4509317

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.

Full text

PDF
3575

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cantor L. N., Morris A. J., Marks P. A., Rifkind R. A. Purification of erythropoietin-responsive cells by immune hemolysis. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1337–1341. doi: 10.1073/pnas.69.6.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chui D. H., Djaldetti M., Marks P. A., Rifkind R. A. Erythropoietin effects on fetal mouse erythroid cells. I. Cell population and hemoglobin synthesis. J Cell Biol. 1971 Dec;51(3):585–595. doi: 10.1083/jcb.51.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gross M., Goldwasser E. On the mechanism of erythropoietin-induced differentiation. V. Characterization of the ribonucleic acid formed as a result of erythropoietin action. Biochemistry. 1969 May;8(5):1795–1805. doi: 10.1021/bi00833a003. [DOI] [PubMed] [Google Scholar]
  4. Jacobs-Lorena M., Baglioni C. Messenger RNA for globin in the postribosomal supernatant of rabbit reticulocytes. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1425–1428. doi: 10.1073/pnas.69.6.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Krantz S. B., Goldwasser E. On the mechanism of erythropoietin-induced differentiation. II. T he effect on RNA synthesis. Biochim Biophys Acta. 1965 Jun 8;103(2):325–332. doi: 10.1016/0005-2787(65)90172-3. [DOI] [PubMed] [Google Scholar]
  6. Lebleu B., Marbaix G., Huez G., Temmerman J., Burny A., Chantrenne H. Characterization of the messenger ribonucleoprotein released from reticulocyte polyribosomes by EDTA treatment. Eur J Biochem. 1971 Mar 11;19(2):264–269. doi: 10.1111/j.1432-1033.1971.tb01313.x. [DOI] [PubMed] [Google Scholar]
  7. Marks P. A., Rifkind R. A. Protein synthesis: its control in erythropoiesis. Science. 1972 Mar 3;175(4025):955–961. doi: 10.1126/science.175.4025.955. [DOI] [PubMed] [Google Scholar]
  8. Metafora S., Terada M., Dow L. W., Marks P. A., Bank A. Increased efficiency of exogenous messenger RNA translation in a Krebs ascites cell lysate. Proc Natl Acad Sci U S A. 1972 May;69(5):1299–1303. doi: 10.1073/pnas.69.5.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Minio F., Howe C., Hsu K. C., Rifkind R. A. Antigen density on differentiating erythroid cells. Nat New Biol. 1972 Jun 7;237(75):187–188. doi: 10.1038/newbio237187a0. [DOI] [PubMed] [Google Scholar]
  10. Nicol A. G., Conkie D., Lanyon W. G., Drewienkiewicz C. E., Williamson R., Paul J. Characteristics of erythropoietin-induced RNA from foetal mouse liver erythropoietic cell cultures and the effects of 5-fluorodeoxyuridine. Biochim Biophys Acta. 1972 Aug 25;277(2):342–353. doi: 10.1016/0005-2787(72)90416-9. [DOI] [PubMed] [Google Scholar]
  11. Stephenson J. R., Axelrad A. A. Separation of erythropoietin-sensitive cells from hemopoietic spleen colony-forming stem cells of mouse fetal liver by unit gravity sedimentation. Blood. 1971 Apr;37(4):417–427. [PubMed] [Google Scholar]
  12. Terada M., Metafora S., Banks J., Dow L. W., Bank A., Marks P. A. Conservation of globin messenger RNA in rabbit reticulocyte monoribosomes after sodium fluoride treatment. Biochem Biophys Res Commun. 1972 May 26;47(4):766–774. doi: 10.1016/0006-291x(72)90558-x. [DOI] [PubMed] [Google Scholar]
  13. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES