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. 1974 Nov;71(11):4551–4555. doi: 10.1073/pnas.71.11.4551

Regulation of Hemoglobin Synthesis in a Murine Erythroblastic Leukemic Cell: The Requirement for Replication to Induce Hemoglobin Synthesis

Patrick R McClintock 1,2, John Papaconstantinou 1,2
PMCID: PMC433925  PMID: 4531000

Abstract

The induction of hemoglobin synthesis by dimethylsulfoxide in cells infected with Friend leukemia virus has been shown to depend on the number of cell divisions after the addition of the inducing agent. In asynchronous cultures the cells have a generation time of 24 hr, and hemoglobin synthesis occurs after 48 hr in the presence of dimethylsulfoxide. Generation times were extended by lowering the serum content of the medium. In 7.5% and 2.5% fetal calf serum, the generation time was increased to 36 hr and 48 hr, respectively, and hemoglobin synthesis began at 72 hr and 96 hr, respectively. In the presence of N6,O2′-dibutyryladenosine 3′:5′-cyclic monophosphate the generation time was extended to 38 hr, and hemoglobin synthesis began after 72 hr in dimethylsulfoxide.

The number of cell doublings, under conditions where the generation time was 24 hr, was regulated by the size of the initial cell concentration. When a low inoculum (1.5 × 105 cells per cm3) was used, the culture was still in log phase after two doublings and hemoglobin appeared in log phase; a moderate initial inoculum (4.2 × 105 cells per cm3) took the culture into stationary phase after two doublings, and hemoglobin synthesis was induced in stationary-phase cells. Heavy inocula (3.7 × 106 cells per cm3) maintained the cells in stationary phase, and under these conditions no induction of hemoglobin synthesis occured. We conclude from these studies that the initiation of hemoglobin synthesis required two rounds of mitosis after treatment with dimethylsulfoxide.

In studies with synchronized cultures of cells infected with Friend leukemia virus, two mitoses were required for the initiation of hemoglobin synthesis. We conclude that two rounds of DNA synthesis and/or mitosis are also required for the dimethylsulfoxide-mediated induction of erythroid differentiation and hemoglobin synthesis in these cells.

Keywords: differentiation, Friend leukemia cells

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Selected References

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

  1. Burger M. M., Bombik B. M., Breckenridge B. M., Sheppard J. R. Growth control and cyclic alterations of cyclic AMP in the cell cycle. Nat New Biol. 1972 Oct 11;239(93):161–163. doi: 10.1038/newbio239161a0. [DOI] [PubMed] [Google Scholar]
  2. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  3. Friend C., Patuleia M. C., De Harven E. Erythrocytic maturation in vitro of murine (Friend) virus-induced leukemic cells. Natl Cancer Inst Monogr. 1966 Sep;22:505–522. [PubMed] [Google Scholar]
  4. Friend C., Scher W., Holland J. G., Sato T. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide. Proc Natl Acad Sci U S A. 1971 Feb;68(2):378–382. doi: 10.1073/pnas.68.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Froehlich J. E., Rachmeler M. Inhibition of cell growth in the G1 phase by adenosine 3', 5'-cyclic monophosphate. J Cell Biol. 1974 Jan;60(1):249–257. doi: 10.1083/jcb.60.1.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Oler A., Iannaccone P. M., Gordon G. B. Suppression of growth of L cells in suspension culture by dibutyrl adenosine 3',5'-monophosphate. In Vitro. 1973 Jul-Aug;9(1):35–38. doi: 10.1007/BF02615987. [DOI] [PubMed] [Google Scholar]
  7. Ostertag W., Crozier T., Kluge N., Melderis H., Dube S. Action of 5-bromodeoxyuridine on the induction of haemoglobin synthesis in mouse leukaemia cells resistant to 5-BUdR. Nat New Biol. 1973 Jun 13;243(128):203–205. doi: 10.1038/newbio243203a0. [DOI] [PubMed] [Google Scholar]
  8. Ostertag W., Melderis H., Steinheider G., Kluge N., Dube S. Synthesis of mouse haemoglobin and globin mRNA in leukaemic cell cultures. Nat New Biol. 1972 Oct 25;239(95):231–234. doi: 10.1038/newbio239231a0. [DOI] [PubMed] [Google Scholar]
  9. Otten J., Johnson G. S., Pastan I. Regulation of cell growth by cyclic adenosine 3',5'-monophosphate. Effect of cell density and agents which alter cell growth on cyclic adenosine 3',5'-monophosphate levels in fibroblasts. J Biol Chem. 1972 Nov 10;247(21):7082–7087. [PubMed] [Google Scholar]
  10. Owens I. S., Vonderhaar B. K., Topper Y. J. Concerning the necessary coupling of development to proliferation of mouse mammary epithelial cells. J Biol Chem. 1973 Jan 25;248(2):472–477. [PubMed] [Google Scholar]
  11. PUCK T. T., CIECIURA S. J., FISHER H. W. Clonal growth in vitro of human cells with fibroblastic morphology; comparison of growth and genetic characteristics of single epithelioid and fibroblast-like cells from a variety of human organs. J Exp Med. 1957 Jul 1;106(1):145–158. doi: 10.1084/jem.106.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Paul J., Hunter J. A. DNA synthesis is essential for increased haemoglobin synthesis in response to erythropoietin. Nature. 1968 Sep 28;219(5161):1362–1363. doi: 10.1038/2191362a0. [DOI] [PubMed] [Google Scholar]
  13. Richelson E. Stimulation of tyrosine hydroxylase activity in an adrenergic clone of mouse neuroblastoma by dibutyryl cyclic AMP. Nat New Biol. 1973 Apr 11;242(119):175–177. doi: 10.1038/newbio242175a0. [DOI] [PubMed] [Google Scholar]
  14. Ross J., Ikawa Y., Leder P. Globin messenger-RNA induction during erythroid differentiation of cultured leukemia cells. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3620–3623. doi: 10.1073/pnas.69.12.3620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ryan W. L., Heidrick M. L. Inhibition of cell growth in vitro by adenosine 3',5'-monophosphate. Science. 1968 Dec 27;162(3861):1484–1485. doi: 10.1126/science.162.3861.1484. [DOI] [PubMed] [Google Scholar]
  16. Sato T., Friend C., De Harven E. Ultrastructural changes in Friend erythroleukemia cells treated with dimethyl sulfoxide. Cancer Res. 1971 Oct;31(10):1402–1417. [PubMed] [Google Scholar]
  17. Scher W., Holland J. G., Friend C. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro. I. Partial purification and identification of hemoglobins. Blood. 1971 Apr;37(4):428–437. [PubMed] [Google Scholar]
  18. Scher W., Preisler H. D., Friend C. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro. 3. Effects of 5-bromo-2'-deoxyuridine, dimethylformamide and dimethylsulfoxide. J Cell Physiol. 1973 Feb;81(1):63–70. doi: 10.1002/jcp.1040810108. [DOI] [PubMed] [Google Scholar]
  19. Thomas D. B., Medley G., Lingwood C. A. Growth inhibition of murine tumor cells, in vitro, by puromycin, ( 6 N)O 2 '-dibutyryl 3',5'-adenosine monophosphate, or adenosine: evidence of commitment for cell division. J Cell Biol. 1973 May;57(2):397–405. doi: 10.1083/jcb.57.2.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Turkington R. W., Majumder G. C., Riddle M. Inhibition of mammary gland differentiation in vitro by 5-bromo-2'-deoxyuridine. J Biol Chem. 1971 Mar 25;246(6):1814–1819. [PubMed] [Google Scholar]
  21. Weintraub H., Campbell G. L., Holtzer H. Identification of a developmental program using bromodeoxyuridine. J Mol Biol. 1972 Sep 28;70(2):337–350. doi: 10.1016/0022-2836(72)90543-8. [DOI] [PubMed] [Google Scholar]

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