Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Jul 1;99(1 Pt 1):306–314. doi: 10.1083/jcb.99.1.306

Differentiation of murine erythroleukemia cells results in the rapid repression of vimentin gene expression

PMCID: PMC2275651  PMID: 6376521

Abstract

We show that vimentin filaments are present in undifferentiated Friend murine erythroleukemia cells, but are lost progressively to undetectable levels by 96 h of dimethyl sulfoxide-mediated differentiation. The amount of newly synthesized cytoskeletal vimentin is decreased dramatically by 24 h of induction, and is paralleled by a rapid loss of vimentin mRNA (approximately 25-fold reduction at 96 h). Hence, disappearance of vimentin filaments in these cells appears to be regulated at the level of vimentin mRNA abundance. On the other hand, the levels of actin synthesis and actin mRNA remain essentially unchanged. The kinetics of vimentin mRNA reduction during dimethyl sulfoxide-mediated differentiation, and the levels of vimentin mRNA observed in the presence of hexamethylene-bisacetamide or hemin as inducers suggest that the cessation of vimentin expression precedes, but may be associated with commitment to terminal differentiation. Our results demonstrate the dynamic regulation of vimentin expression in mammalian erythropoiesis.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Voloch Z., Bastos R., Levy S. Biosynthesis and stability of globin mRNA in cultured erythroleukemic Friend cells. Cell. 1976 Aug;8(4):495–503. doi: 10.1016/0092-8674(76)90217-8. [DOI] [PubMed] [Google Scholar]
  3. Blikstad I., Lazarides E. Vimentin filaments are assembled from a soluble precursor in avian erythroid cells. J Cell Biol. 1983 Jun;96(6):1803–1808. doi: 10.1083/jcb.96.6.1803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blikstad I., Nelson W. J., Moon R. T., Lazarides E. Synthesis and assembly of spectrin during avian erythropoiesis: stoichiometric assembly but unequal synthesis of alpha and beta spectrin. Cell. 1983 Apr;32(4):1081–1091. doi: 10.1016/0092-8674(83)90292-1. [DOI] [PubMed] [Google Scholar]
  5. Capetanaki Y. G., Ngai J., Flytzanis C. N., Lazarides E. Tissue-specific expression of two mRNA species transcribed from a single vimentin gene. Cell. 1983 Dec;35(2 Pt 1):411–420. doi: 10.1016/0092-8674(83)90174-5. [DOI] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Dellagi K., Vainchenker W., Vinci G., Paulin D., Brouet J. C. Alteration of vimentin intermediate filament expression during differentiation of human hemopoietic cells. EMBO J. 1983;2(9):1509–1514. doi: 10.1002/j.1460-2075.1983.tb01615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dodemont H. J., Soriano P., Quax W. J., Ramaekers F., Lenstra J. A., Groenen M. A., Bernardi G., Bloemendal H. The genes coding for the cytoskeletal proteins actin and vimentin in warm-blooded vertebrates. EMBO J. 1982;1(2):167–171. doi: 10.1002/j.1460-2075.1982.tb01142.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ebert P. S., Ikawa Y. Induction of delta-aminolevulinic acid synthetase during erythroid differentiation of cultured leukemia cells. Proc Soc Exp Biol Med. 1974 Jun;146(2):601–604. doi: 10.3181/00379727-146-38155. [DOI] [PubMed] [Google Scholar]
  10. Fibach E., Reuben R. C., Rifkind R. A., Marks P. A. Effect of hexamethylene bisacetamide on the commitment to differentiation of murine erythroleukemia cells. Cancer Res. 1977 Feb;37(2):440–444. [PubMed] [Google Scholar]
  11. Friedman E. A., Schildkraut C. L. Terminal differentiation in cultured Friend erythroleukemia cells. Cell. 1977 Dec;12(4):901–913. doi: 10.1016/0092-8674(77)90154-4. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Gard D. L., Lazarides E. The synthesis and distribution of desmin and vimentin during myogenesis in vitro. Cell. 1980 Jan;19(1):263–275. doi: 10.1016/0092-8674(80)90408-0. [DOI] [PubMed] [Google Scholar]
  15. Goldberg D. A. Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5794–5798. doi: 10.1073/pnas.77.10.5794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Granger B. L., Lazarides E. Desmin and vimentin coexist at the periphery of the myofibril Z disc. Cell. 1979 Dec;18(4):1053–1063. doi: 10.1016/0092-8674(79)90218-6. [DOI] [PubMed] [Google Scholar]
  17. Granger B. L., Lazarides E. Structural associations of synemin and vimentin filaments in avian erythrocytes revealed by immunoelectron microscopy. Cell. 1982 Aug;30(1):263–275. doi: 10.1016/0092-8674(82)90032-0. [DOI] [PubMed] [Google Scholar]
  18. Granger B. L., Repasky E. A., Lazarides E. Synemin and vimentin are components of intermediate filaments in avian erythrocytes. J Cell Biol. 1982 Feb;92(2):299–312. doi: 10.1083/jcb.92.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Groudine M., Peretz M., Weintraub H. Transcriptional regulation of hemoglobin switching in chicken embryos. Mol Cell Biol. 1981 Mar;1(3):281–288. doi: 10.1128/mcb.1.3.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gusella J. F., Weil S. C., Tsiftsoglou A. S., Volloch V., Neumann J. R., Keys C., Housman D. E. Hemin does not cause commitment of murine erythroleukemia (MEL) cells to terminal differentiation. Blood. 1980 Sep;56(3):481–487. [PubMed] [Google Scholar]
  21. Gusella J., Geller R., Clarke B., Weeks V., Housman D. Commitment to erythroid differentiation by friend erythroleukemia cells: a stochastic analysis. Cell. 1976 Oct;9(2):221–229. doi: 10.1016/0092-8674(76)90113-6. [DOI] [PubMed] [Google Scholar]
  22. Hubbard B. D., Lazarides E. Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments. J Cell Biol. 1979 Jan;80(1):166–182. doi: 10.1083/jcb.80.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Keppel F., Allet B., Eisen H. Appearance of a chromatin protein during the erythroid differentiation of Friend virus-transformed cells. Proc Natl Acad Sci U S A. 1977 Feb;74(2):653–656. doi: 10.1073/pnas.74.2.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Konkel D. A., Tilghman S. M., Leder P. The sequence of the chromosomal mouse beta-globin major gene: homologies in capping, splicing and poly(A) sites. Cell. 1978 Dec;15(4):1125–1132. doi: 10.1016/0092-8674(78)90040-5. [DOI] [PubMed] [Google Scholar]
  25. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  26. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  27. Leder A., Leder P. Butyric acid, a potent inducer of erythroid differentiation in cultured erythroleukemic cells. Cell. 1975 Jul;5(3):319–322. doi: 10.1016/0092-8674(75)90107-5. [DOI] [PubMed] [Google Scholar]
  28. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  29. Levy J., Terada M., Rifkind R. A., Marks P. A. Induction of erythroid differentiation by dimethylsulfoxide in cells infected with Friend virus: relationship to the cell cycle. Proc Natl Acad Sci U S A. 1975 Jan;72(1):28–32. doi: 10.1073/pnas.72.1.28. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lowenhaupt K., Lingrel J. B. A change in the stability of globin mRNA during the induction of murine erythroleukemia cells. Cell. 1978 Jun;14(2):337–344. doi: 10.1016/0092-8674(78)90119-8. [DOI] [PubMed] [Google Scholar]
  31. Lowenhaupt K., Lingrel J. B. Synthesis and turnover of globin mRNA in murine erythroleukemia cells induced with hemin. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5173–5177. doi: 10.1073/pnas.76.10.5173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lowenhaupt K., Trent C., Lingrel J. B. Mechanisms for accumulation of globin mRNA during dimethyl sulfoxide induction of murine erythroleukemia cells: synthesis of precursors and mature mRNA. Dev Biol. 1978 Apr;63(2):441–454. doi: 10.1016/0012-1606(78)90148-3. [DOI] [PubMed] [Google Scholar]
  33. Marks P. A., Rifkind R. A. Erythroleukemic differentiation. Annu Rev Biochem. 1978;47:419–448. doi: 10.1146/annurev.bi.47.070178.002223. [DOI] [PubMed] [Google Scholar]
  34. McClintock P. R., Papaconstantinou J. Regulation of hemoglobin synthesis in a murine erythroblastic leukemic cell: the requirement for replication to induce hemoglobin synthesis. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4551–4555. doi: 10.1073/pnas.71.11.4551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  36. McTavish C. F., Nelson W. J., Traub P. The turnover of vimentin in Ehrlich ascites tumour cells. FEBS Lett. 1983 Apr 18;154(2):251–256. doi: 10.1016/0014-5793(83)80159-8. [DOI] [PubMed] [Google Scholar]
  37. Moon R. T., Lazarides E. Synthesis and post-translational assembly of intermediate filaments in avian erythroid cells: vimentin assembly limits the rate of synemin assembly. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5495–5499. doi: 10.1073/pnas.80.18.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nelson W. J., Vorgias C. E., Traub P. A rapid method for the large scale purification of the intermediate filament protein vimentin by single-stranded DNA-cellulose affinity chromatography. Biochem Biophys Res Commun. 1982 Jun 30;106(4):1141–1147. doi: 10.1016/0006-291x(82)91231-1. [DOI] [PubMed] [Google Scholar]
  39. Nudel U., Salmon J., Fibach E., Terada M., Rifkind R., Marks P. A., Bank A. Accumulation of alpha- and beta-globin messenger RNAs in mouse erythroleukemia cells. Cell. 1977 Oct;12(2):463–469. doi: 10.1016/0092-8674(77)90122-2. [DOI] [PubMed] [Google Scholar]
  40. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  41. Orkin S. H., Harosi F. I., Leder P. Differentiation in erythroleukemic cells and their somatic hybrids. Proc Natl Acad Sci U S A. 1975 Jan;72(1):98–102. doi: 10.1073/pnas.72.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Orkin S. H., Swerdlow P. S. Globin RNA synthesis in vitro by isolated erythroleukemic cell nuclei: direct evidence for increased transcription during erythroid differentiation. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2475–2479. doi: 10.1073/pnas.74.6.2475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Quax W., Egberts W. V., Hendriks W., Quax-Jeuken Y., Bloemendal H. The structure of the vimentin gene. Cell. 1983 Nov;35(1):215–223. doi: 10.1016/0092-8674(83)90224-6. [DOI] [PubMed] [Google Scholar]
  44. Reem G. H., Friend C. Purine metabolism in murine virus-induced erythroleukemic cells during differentiation in vitro. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1630–1634. doi: 10.1073/pnas.72.4.1630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Repasky E. A., Eckert B. S. A reevaluation of the process of enucleation in mammalian erythroid cells. Prog Clin Biol Res. 1981;55:679–692. [PubMed] [Google Scholar]
  46. Reuben R. C., Wife R. L., Breslow R., Rifkind R. A., Marks P. A. A new group of potent inducers of differentiation in murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1976 Mar;73(3):862–866. doi: 10.1073/pnas.73.3.862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Ross J., Sautner D. Induction of globin mRNA accumulation by hemin in cultured erythroleukemic cells. Cell. 1976 Aug;8(4):513–520. doi: 10.1016/0092-8674(76)90219-1. [DOI] [PubMed] [Google Scholar]
  50. Rovera G., Surrey S. Use of resistant or hypersensitive variant clones of Friend cells in analysis of mode of action of inducers. Cancer Res. 1978 Nov;38(11 Pt 1):3737–3744. [PubMed] [Google Scholar]
  51. Sabban E. L., Sabatini D. D., Marchesi V. T., Adesnik M. Biosynthesis of erythrocyte membrane protein band 3 in DMSO-induced Friend erythroleukemia cells. J Cell Physiol. 1980 Aug;104(2):261–268. doi: 10.1002/jcp.1041040217. [DOI] [PubMed] [Google Scholar]
  52. Sassa S. Sequential induction of heme pathway enzymes during erythroid differentiation of mouse Friend leukemia virus-infected cells. J Exp Med. 1976 Feb 1;143(2):305–315. doi: 10.1084/jem.143.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Scheller R. H., McAllister L. B., Crain W. R., Jr, Durica D. S., Posakony J. W., Thomas T. L., Britten R. J., Davidson E. H. Organization and expression of multiple actin genes in the sea urchin. Mol Cell Biol. 1981 Jul;1(7):609–628. doi: 10.1128/mcb.1.7.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Shaul Y., Ginzburg I., Aviv H. Preferential transcription and nuclear transport of globin gene sequences, as control steps leading to final differentiation of murine erythroleukemic cells. Eur J Biochem. 1982 Nov 15;128(2-3):637–642. doi: 10.1111/j.1432-1033.1982.tb07011.x. [DOI] [PubMed] [Google Scholar]
  55. Sherton C. C., Kabat D. Changes in RNA and protein metabolism preceding onset of hemoglobin synthesis in cultured Friend leukemia cells. Dev Biol. 1976 Jan;48(1):118–131. doi: 10.1016/0012-1606(76)90051-8. [DOI] [PubMed] [Google Scholar]
  56. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  57. Tapscott S. J., Bennett G. S., Toyama Y., Kleinbart F., Holtzer H. Intermediate filament proteins in the developing chick spinal cord. Dev Biol. 1981 Aug;86(1):40–54. doi: 10.1016/0012-1606(81)90313-4. [DOI] [PubMed] [Google Scholar]
  58. Volloch V., Housman D. Stability of globin mRNA in terminally differentiating murine erythroleukemia cells. Cell. 1981 Feb;23(2):509–514. doi: 10.1016/0092-8674(81)90146-x. [DOI] [PubMed] [Google Scholar]
  59. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Yen S. H., Fields K. L. Antibodies to neurofilament, glial filament, and fibroblast intermediate filament proteins bind to different cell types of the nervous system. J Cell Biol. 1981 Jan;88(1):115–126. doi: 10.1083/jcb.88.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES