Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1987 Jan;7(1):365–370. doi: 10.1128/mcb.7.1.365

Rearrangement and expression of erythropoietin genes in transformed mouse cells.

J McDonald, N Beru, E Goldwasser
PMCID: PMC365077  PMID: 3561395

Abstract

The erythroleukemia cell line IW32, derived by transformation with the Friend murine leukemia virus, has been shown previously to produce erythropoietin (EPO) constitutively. Here we demonstrate that, in addition to the normal mouse EPO locus, this cell line has another EPO locus which has undergone rearrangement and amplification. Both loci were cloned, and the rearrangement breakpoint of the second EPO locus was located within a 1.1-kilobase region upstream of an otherwise apparently normal EPO gene. There are no viral sequences present in the immediate vicinity of the rearranged EPO gene. DNase I digestion studies suggest that the rearranged gene is in a region where the chromatin is more sensitive to DNase hydrolysis than is the site of the normal gene. We conclude, tentatively, that the rearranged EPO locus is probably the transcriptionally active one and that either proviral sequences are acting at a distance to activate the EPO gene or the rearrangement itself has served to activate the gene.

Full text

PDF
365

Images in this article

366Image
on p.366
367Image
on p.367
367Image
on p.367
368Image
on p.368
368Image
on p.368
368Image
on p.368

Selected References

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

  1. Affara N., Fleming J., Goldfarb P. S., Black E., Thiele B., Harrison P. R. Analysis of chromatin changes associated with the expression of globin and non-globin genes in cell hybrids between erythroid and other cells. Nucleic Acids Res. 1985 Aug 12;13(15):5629–5644. doi: 10.1093/nar/13.15.5629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beru N., McDonald J., Lacombe C., Goldwasser E. Expression of the erythropoietin gene. Mol Cell Biol. 1986 Jul;6(7):2571–2575. doi: 10.1128/mcb.6.7.2571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bondurant M. C., Koury M. J. Anemia induces accumulation of erythropoietin mRNA in the kidney and liver. Mol Cell Biol. 1986 Jul;6(7):2731–2733. doi: 10.1128/mcb.6.7.2731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burch J. B., Weintraub H. Temporal order of chromatin structural changes associated with activation of the major chicken vitellogenin gene. Cell. 1983 May;33(1):65–76. doi: 10.1016/0092-8674(83)90335-5. [DOI] [PubMed] [Google Scholar]
  6. Choppin J., Casadevall N., Lacombe C., Wendling F., Goldwasser E., Berger R., Tambourin P., Varet B. Production of erythropoietin by cloned malignant murine erythroid cells. Exp Hematol. 1985 Aug;13(7):610–615. [PubMed] [Google Scholar]
  7. Choppin J., Lacombe C., Casadevall N., Muller O., Tambourin P., Varet B. Characterization of erythropoietin produced by IW32 murine erythroleukemia cells. Blood. 1984 Aug;64(2):341–347. [PubMed] [Google Scholar]
  8. Frischauf A. M., Lehrach H., Poustka A., Murray N. Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983 Nov 15;170(4):827–842. doi: 10.1016/s0022-2836(83)80190-9. [DOI] [PubMed] [Google Scholar]
  9. Fung Y. K., Lewis W. G., Crittenden L. B., Kung H. J. Activation of the cellular oncogene c-erbB by LTR insertion: molecular basis for induction of erythroblastosis by avian leukosis virus. Cell. 1983 Jun;33(2):357–368. doi: 10.1016/0092-8674(83)90417-8. [DOI] [PubMed] [Google Scholar]
  10. Goldwasser E., Eliason J. F., Sikkema D. An assay for erythropoietin in vitro at the milliunit level. Endocrinology. 1975 Aug;97(2):315–323. doi: 10.1210/endo-97-2-315. [DOI] [PubMed] [Google Scholar]
  11. Groudine M., Weintraub H. Propagation of globin DNAase I-hypersensitive sites in absence of factors required for induction: a possible mechanism for determination. Cell. 1982 Aug;30(1):131–139. doi: 10.1016/0092-8674(82)90019-8. [DOI] [PubMed] [Google Scholar]
  12. Hayward W. S., Neel B. G., Astrin S. M. Activation of a cellular onc gene by promoter insertion in ALV-induced lymphoid leukosis. Nature. 1981 Apr 9;290(5806):475–480. doi: 10.1038/290475a0. [DOI] [PubMed] [Google Scholar]
  13. Hillova J., Hill M., Belehradek J., Mariage-Samson R., Marx M. Amplification of the provirus region in Rous sarcoma virus-transformed Chinese hamster cells and segregation of the amplified copies in somatic cell hybrids. Virology. 1983 Jul 15;128(1):1–15. doi: 10.1016/0042-6822(83)90314-8. [DOI] [PubMed] [Google Scholar]
  14. Hu N., Messing J. The making of strand-specific M13 probes. Gene. 1982 Mar;17(3):271–277. doi: 10.1016/0378-1119(82)90143-3. [DOI] [PubMed] [Google Scholar]
  15. Ishimoto A., Adachi A., Sakai K., Yorifuji T., Tsuruta S. Rapid emergence of mink cell focus-forming (MCF) virus in various mice infected with NB-tropic friend virus. Virology. 1981 Sep;113(2):644–655. doi: 10.1016/0042-6822(81)90193-8. [DOI] [PubMed] [Google Scholar]
  16. JACOBSON L. O., GOLDWASSER E., FRIED W., PLZAK L. Role of the kidney in erythropoiesis. Nature. 1957 Mar 23;179(4560):633–634. doi: 10.1038/179633a0. [DOI] [PubMed] [Google Scholar]
  17. Jacobs K., Shoemaker C., Rudersdorf R., Neill S. D., Kaufman R. J., Mufson A., Seehra J., Jones S. S., Hewick R., Fritsch E. F. Isolation and characterization of genomic and cDNA clones of human erythropoietin. 1985 Feb 28-Mar 6Nature. 313(6005):806–810. doi: 10.1038/313806a0. [DOI] [PubMed] [Google Scholar]
  18. Leder P., Battey J., Lenoir G., Moulding C., Murphy W., Potter H., Stewart T., Taub R. Translocations among antibody genes in human cancer. Science. 1983 Nov 18;222(4625):765–771. doi: 10.1126/science.6356357. [DOI] [PubMed] [Google Scholar]
  19. Levy L. S., Gardner M. B., Casey J. W. Isolation of a feline leukaemia provirus containing the oncogene myc from a feline lymphosarcoma. 1984 Apr 26-May 2Nature. 308(5962):853–856. doi: 10.1038/308853a0. [DOI] [PubMed] [Google Scholar]
  20. Lin F. K., Suggs S., Lin C. H., Browne J. K., Smalling R., Egrie J. C., Chen K. K., Fox G. M., Martin F., Stabinsky Z. Cloning and expression of the human erythropoietin gene. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7580–7584. doi: 10.1073/pnas.82.22.7580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McDonald J. D., Lin F. K., Goldwasser E. Cloning, sequencing, and evolutionary analysis of the mouse erythropoietin gene. Mol Cell Biol. 1986 Mar;6(3):842–848. doi: 10.1128/mcb.6.3.842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mullins J. I., Brody D. S., Binari R. C., Jr, Cotter S. M. Viral transduction of c-myc gene in naturally occurring feline leukaemias. 1984 Apr 26-May 2Nature. 308(5962):856–858. doi: 10.1038/308856a0. [DOI] [PubMed] [Google Scholar]
  23. Neil J. C., Hughes D., McFarlane R., Wilkie N. M., Onions D. E., Lees G., Jarrett O. Transduction and rearrangement of the myc gene by feline leukaemia virus in naturally occurring T-cell leukaemias. 1984 Apr 26-May 2Nature. 308(5962):814–820. doi: 10.1038/308814a0. [DOI] [PubMed] [Google Scholar]
  24. Noori-Daloii M. R., Swift R. A., Kung H. J., Crittenden L. B., Witter R. L. Specific integration of REV proviruses in avian bursal lymphomas. Nature. 1981 Dec 10;294(5841):574–576. doi: 10.1038/294574a0. [DOI] [PubMed] [Google Scholar]
  25. Nusse R., Varmus H. E. Many tumors induced by the mouse mammary tumor virus contain a provirus integrated in the same region of the host genome. Cell. 1982 Nov;31(1):99–109. doi: 10.1016/0092-8674(82)90409-3. [DOI] [PubMed] [Google Scholar]
  26. Oliff A. I., Hager G. L., Chang E. H., Scolnick E. M., Chan H. W., Lowy D. R. Transfection of molecularly cloned Friend murine leukemia virus DNA yields a highly leukemogenic helper-independent type C virus. J Virol. 1980 Jan;33(1):475–486. doi: 10.1128/jvi.33.1.475-486.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Parsons J. T., Coffin J. M., Haroz R. K., Bromley P. A., Weissmann C. Quantitative determination and location of newly synthesized virus-specific ribonucleic acid in chicken cells infected with Rous sarcoma virus. J Virol. 1973 May;11(5):761–774. doi: 10.1128/jvi.11.5.761-774.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Payne G. S., Courtneidge S. A., Crittenden L. B., Fadly A. M., Bishop J. M., Varmus H. E. Analysis of avian leukosis virus DNA and RNA in bursal tumours: viral gene expression is not required for maintenance of the tumor state. Cell. 1981 Feb;23(2):311–322. doi: 10.1016/0092-8674(81)90127-6. [DOI] [PubMed] [Google Scholar]
  29. Peschle C., Mavilio F., Sposi N. M., Giampaolo A., Caré A., Bottero L., Bruno M., Mastroberardino G., Gastaldi R., Testa M. G. Translocation and rearrangement of c-myc into immunoglobulin alpha heavy chain locus in primary cells from acute lymphocytic leukemia. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5514–5518. doi: 10.1073/pnas.81.17.5514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Robertson M. Paradox and paradigm: the message and meaning of myc. Nature. 1983 Dec 22;306(5945):733–736. doi: 10.1038/306733a0. [DOI] [PubMed] [Google Scholar]
  32. Sherwood J. B., Goldwasser E. A radioimmunoassay for erythropoietin. Blood. 1979 Oct;54(4):885–893. [PubMed] [Google Scholar]
  33. Shiraishi A., Nakanishi Y., Sekimizu K., Natori S. Analysis of the DNase I-hypersensitive site of a developmentally regulated 25-kDa protein gene of Sarcophaga peregrina. J Biol Chem. 1986 Jan 15;261(2):940–943. [PubMed] [Google Scholar]
  34. Shoemaker C. B., Mitsock L. D. Murine erythropoietin gene: cloning, expression, and human gene homology. Mol Cell Biol. 1986 Mar;6(3):849–858. doi: 10.1128/mcb.6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Stark G. R., Wahl G. M. Gene amplification. Annu Rev Biochem. 1984;53:447–491. doi: 10.1146/annurev.bi.53.070184.002311. [DOI] [PubMed] [Google Scholar]
  37. Tambourin P., Casadevall N., Choppin J., Lacombe C., Heard J. M., Fichelson S., Wendling F., Varet B. Production of erythropoietin-like activity by a murine erythroleukemia cell line. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6269–6273. doi: 10.1073/pnas.80.20.6269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]
  39. Zucali J. R., Stevens V., Mirand E. A. In vitro production of erythropoietin by mouse fetal liver. Blood. 1975 Jul;46(1):85–90. [PubMed] [Google Scholar]
  40. van Ooyen A., Nusse R. Structure and nucleotide sequence of the putative mammary oncogene int-1; proviral insertions leave the protein-encoding domain intact. Cell. 1984 Nov;39(1):233–240. doi: 10.1016/0092-8674(84)90209-5. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES