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. 1986 May 12;14(9):3641–3658. doi: 10.1093/nar/14.9.3641

Cloning and expression of the rat interleukin-3 gene.

D R Cohen, A J Hapel, I G Young
PMCID: PMC339805  PMID: 3086845

Abstract

Genomic clones carrying the rat interleukin-3 (IL-3) gene have been isolated and the nucleotide sequence of the gene determined. Alignment of this sequence with that of the mouse IL-3 gene has allowed the structure of the rat IL-3 gene to be deduced. The intron-exon boundaries are conserved and extensive nucleotide homology (approx 90%) is present in the 5' flanking region and the portion of the gene coding for the signal peptide. Several proposed regulatory sequences are conserved and an analogous element to the tandem repeat in intron 2 of the mouse gene is also present. The predicted amino acid sequence for mature rat IL-3 shows surprisingly low homology (54%) with its murine counterpart, although all four cysteine residues are conserved. The rat IL-3 gene was expressed in monkey COS-1 cells and colony assays established that rat IL-3 is a multi-lineage haemopoietic growth regulator. There was little cross-reactivity of the respective IL-3 species on mouse and rat bone marrow cells suggesting that rat IL-3, in concert with its receptor, has evolved significantly away from the mouse IL-3/receptor system.

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Selected References

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

  1. Abbud-Filho M., Kupiec-Weglinski J. W., Araujo J. L., Heidecke C. D., Tilney N. L., Strom T. B. Cyclosporine therapy of rat heart allograft recipients and release of interleukins (IL 1, IL 2, IL 3): a role for IL 3 in graft tolerance? J Immunol. 1984 Nov;133(5):2582–2586. [PubMed] [Google Scholar]
  2. Alonso A., Kühn B., Fischer J. An unusual accumulation of repetitive sequences in the rat genome. Gene. 1983 Dec;26(2-3):303–306. doi: 10.1016/0378-1119(83)90202-0. [DOI] [PubMed] [Google Scholar]
  3. Bell G. I., Selby M. J., Rutter W. J. The highly polymorphic region near the human insulin gene is composed of simple tandemly repeating sequences. Nature. 1982 Jan 7;295(5844):31–35. doi: 10.1038/295031a0. [DOI] [PubMed] [Google Scholar]
  4. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Burgess A. W., Camakaris J., Metcalf D. Purification and properties of colony-stimulating factor from mouse lung-conditioned medium. J Biol Chem. 1977 Mar 25;252(6):1998–2003. [PubMed] [Google Scholar]
  7. Burgess A. W., Metcalf D., Russell S. H., Nicola N. A. Granulocyte/macrophage-, megakaryocyte-, eosinophil- and erythroid-colony-stimulating factors produced by mouse spleen cells. Biochem J. 1980 Feb 1;185(2):301–314. doi: 10.1042/bj1850301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Campbell H. D., Ymer S., Fung M. C., Young I. G. Cloning and nucleotide sequence of the murine interleukin-3 gene. Eur J Biochem. 1985 Jul 15;150(2):297–304. doi: 10.1111/j.1432-1033.1985.tb09020.x. [DOI] [PubMed] [Google Scholar]
  9. Cantrell M. A., Anderson D., Cerretti D. P., Price V., McKereghan K., Tushinski R. J., Mochizuki D. Y., Larsen A., Grabstein K., Gillis S. Cloning, sequence, and expression of a human granulocyte/macrophage colony-stimulating factor. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6250–6254. doi: 10.1073/pnas.82.18.6250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carlson D. P., Ross J. Human beta-globin promoter and coding sequences transcribed by RNA polymerase III. Cell. 1983 Oct;34(3):857–864. doi: 10.1016/0092-8674(83)90543-3. [DOI] [PubMed] [Google Scholar]
  11. Clark-Lewis I., Kent S. B., Schrader J. W. Purification to apparent homogeneity of a factor stimulating the growth of multiple lineages of hemopoietic cells. J Biol Chem. 1984 Jun 25;259(12):7488–7494. [PubMed] [Google Scholar]
  12. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  13. Conlon P. J., Luk K. H., Park L. S., March C. J., Hopp T. P., Urdal D. L. Generation of anti-peptide monoclonal antibodies which recognize mature CSF-2 alpha (IL 3) protein. J Immunol. 1985 Jul;135(1):328–332. [PubMed] [Google Scholar]
  14. Crowe W. E., Battisto J. R., Smith R. N. The autologous mixed lymphocyte reaction is decreased in Freund's adjuvant-injected rats of arthritis-susceptible and -insusceptible strains. Arthritis Rheum. 1985 May;28(5):537–541. doi: 10.1002/art.1780280512. [DOI] [PubMed] [Google Scholar]
  15. Deininger P. L. Random subcloning of sonicated DNA: application to shotgun DNA sequence analysis. Anal Biochem. 1983 Feb 15;129(1):216–223. doi: 10.1016/0003-2697(83)90072-6. [DOI] [PubMed] [Google Scholar]
  16. Dijkema R., van der Meide P. H., Pouwels P. H., Caspers M., Dubbeld M., Schellekens H. Cloning and expression of the chromosomal immune interferon gene of the rat. EMBO J. 1985 Mar;4(3):761–767. doi: 10.1002/j.1460-2075.1985.tb03694.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Duckworth M. L., Gait M. J., Goelet P., Hong G. F., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. doi: 10.1093/nar/9.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. EBBE S., STOHLMAN F., Jr MEGAKARYOCYTOPOIESIS IN THE RAT. Blood. 1965 Jul;26:20–35. [PubMed] [Google Scholar]
  19. Fujita T., Takaoka C., Matsui H., Taniguchi T. Structure of the human interleukin 2 gene. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7437–7441. doi: 10.1073/pnas.80.24.7437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fung M. C., Hapel A. J., Ymer S., Cohen D. R., Johnson R. M., Campbell H. D., Young I. G. Molecular cloning of cDNA for murine interleukin-3. Nature. 1984 Jan 19;307(5948):233–237. doi: 10.1038/307233a0. [DOI] [PubMed] [Google Scholar]
  21. Garoff H., Ansorge W. Improvements of DNA sequencing gels. Anal Biochem. 1981 Aug;115(2):450–457. doi: 10.1016/0003-2697(81)90031-2. [DOI] [PubMed] [Google Scholar]
  22. Goodbourn S. E., Higgs D. R., Clegg J. B., Weatherall D. J. Molecular basis of length polymorphism in the human zeta-globin gene complex. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5022–5026. doi: 10.1073/pnas.80.16.5022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gough N. M., Gough J., Metcalf D., Kelso A., Grail D., Nicola N. A., Burgess A. W., Dunn A. R. Molecular cloning of cDNA encoding a murine haematopoietic growth regulator, granulocyte-macrophage colony stimulating factor. 1984 Jun 28-Jul 4Nature. 309(5971):763–767. doi: 10.1038/309763a0. [DOI] [PubMed] [Google Scholar]
  24. Gray P. W., Goeddel D. V. Cloning and expression of murine immune interferon cDNA. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5842–5846. doi: 10.1073/pnas.80.19.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gray P. W., Goeddel D. V. Structure of the human immune interferon gene. Nature. 1982 Aug 26;298(5877):859–863. doi: 10.1038/298859a0. [DOI] [PubMed] [Google Scholar]
  26. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  27. Haig D. M., McKee T. A., Jarrett E. E., Woodbury R., Miller H. R. Generation of mucosal mast cells is stimulated in vitro by factors derived from T cells of helminth-infected rats. Nature. 1982 Nov 11;300(5888):188–190. doi: 10.1038/300188a0. [DOI] [PubMed] [Google Scholar]
  28. Haig D. M., McMenamin C., Gunneberg C., Woodbury R., Jarrett E. E. Stimulation of mucosal mast cell growth in normal and nude rat bone marrow cultures. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4499–4503. doi: 10.1073/pnas.80.14.4499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  30. Hapel A. J., Fung M. C., Johnson R. M., Young I. G., Johnson G., Metcalf D. Biologic properties of molecularly cloned and expressed murine interleukin-3. Blood. 1985 Jun;65(6):1453–1459. [PubMed] [Google Scholar]
  31. Howard M., Burgess A., McPhee D., Metcalf D. T-cell hybridoma secreting hemopoietic regulatory molecules: granulocyte-macrophage and eosinophil colony-stimulating factors. Cell. 1979 Dec;18(4):993–999. doi: 10.1016/0092-8674(79)90211-3. [DOI] [PubMed] [Google Scholar]
  32. Ihle J. N., Lee J. C., Rebar L. T cell recognition of Moloney leukemia virus proteins. III. T cell proliferative responses against gp70 are associated with the production of a lymphokine inducing 20 alpha-hydroxysteroid dehydrogenase in splenic lymphocytes. J Immunol. 1981 Dec;127(6):2565–2570. [PubMed] [Google Scholar]
  33. Ihle J. N., Rebar L., Keller J., Lee J. C., Hapel A. J. Interleukin 3: possible roles in the regulation of lymphocyte differentiation and growth. Immunol Rev. 1982;63:5–32. doi: 10.1111/j.1600-065x.1982.tb00409.x. [DOI] [PubMed] [Google Scholar]
  34. Jeffreys A. J., Wilson V., Thein S. L. Hypervariable 'minisatellite' regions in human DNA. Nature. 1985 Mar 7;314(6006):67–73. doi: 10.1038/314067a0. [DOI] [PubMed] [Google Scholar]
  35. Kalb V. F., Glasser S., King D., Lingrel J. B. A cluster of repetitive elements within a 700 base pair region in the mouse genome. Nucleic Acids Res. 1983 Apr 11;11(7):2177–2184. doi: 10.1093/nar/11.7.2177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Kashima N., Nishi-Takaoka C., Fujita T., Taki S., Yamada G., Hamuro J., Taniguchi T. Unique structure of murine interleukin-2 as deduced from cloned cDNAs. 1985 Jan 31-Feb 6Nature. 313(6001):402–404. doi: 10.1038/313402a0. [DOI] [PubMed] [Google Scholar]
  37. Kramerov D. A., Grigoryan A. A., Ryskov A. P., Georgiev G. P. Long double-stranded sequences (dsRNA-B) of nuclear pre-mRNA consist of a few highly abundant classes of sequences: evidence from DNA cloning experiments. Nucleic Acids Res. 1979 Feb;6(2):697–713. doi: 10.1093/nar/6.2.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Kramerov D. A., Tillib S. V., Ryskov A. P., Georgiev G. P. Nucleotide sequence of small polyadenylated B2 RNA. Nucleic Acids Res. 1985 Sep 25;13(18):6423–6437. doi: 10.1093/nar/13.18.6423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lafferty K. J., Prowse S. J., Al-Adra A., Warren H. S., Vasalli J., Reich E. An improved assay for interleukin 2 (lymphocyte growth factor) produced by mitogen-activated lymphocytes. Aust J Exp Biol Med Sci. 1980 Dec;58(6):533–544. doi: 10.1038/icb.1980.55. [DOI] [PubMed] [Google Scholar]
  40. Lowry R. P., Forbes R. D. Immune mechanisms in organ allograft rejection: IV. An overview of adoptive transfer studies on the pathogenesis of homograft rejection in the inbred rat. Transplant Proc. 1985 Aug;17(4):1963–1965. [PubMed] [Google Scholar]
  41. McMenamin C., Jarrett E. E., Sanderson A. Surface phenotype of T cells producing growth of mucosal mast cells in normal rat bone marrow culture. Immunology. 1985 Jul;55(3):399–403. [PMC free article] [PubMed] [Google Scholar]
  42. Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
  43. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  44. Miyatake S., Yokota T., Lee F., Arai K. Structure of the chromosomal gene for murine interleukin 3. Proc Natl Acad Sci U S A. 1985 Jan;82(2):316–320. doi: 10.1073/pnas.82.2.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Montell C., Fisher E. F., Caruthers M. H., Berk A. J. Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3' consensus sequence AAUAAA. Nature. 1983 Oct 13;305(5935):600–605. doi: 10.1038/305600a0. [DOI] [PubMed] [Google Scholar]
  46. Nevins J. R. The pathway of eukaryotic mRNA formation. Annu Rev Biochem. 1983;52:441–466. doi: 10.1146/annurev.bi.52.070183.002301. [DOI] [PubMed] [Google Scholar]
  47. Parker B. A., Stark G. R. Regulation of simian virus 40 transcription: sensitive analysis of the RNA species present early in infections by virus or viral DNA. J Virol. 1979 Aug;31(2):360–369. doi: 10.1128/jvi.31.2.360-369.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rolovic Z., Jovanovic T., Stankovic Z., Marinkovic N. Abnormal megakaryocytopoiesis in the Belgrade laboratory rat. Blood. 1985 Jan;65(1):60–64. [PubMed] [Google Scholar]
  49. Rosenthal N., Kress M., Gruss P., Khoury G. BK viral enhancer element and a human cellular homolog. Science. 1983 Nov 18;222(4625):749–755. doi: 10.1126/science.6314501. [DOI] [PubMed] [Google Scholar]
  50. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol. 1982 Dec 25;162(4):729–773. doi: 10.1016/0022-2836(82)90546-0. [DOI] [PubMed] [Google Scholar]
  51. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Scalenghe F., Turco E., Edström J. E., Pirrotta V., Melli M. Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma. 1981;82(2):205–216. doi: 10.1007/BF00286105. [DOI] [PubMed] [Google Scholar]
  53. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  54. Staden R. A computer program to enter DNA gel reading data into a computer. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):499–503. doi: 10.1093/nar/12.1part2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Staden R. A new computer method for the storage and manipulation of DNA gel reading data. Nucleic Acids Res. 1980 Aug 25;8(16):3673–3694. doi: 10.1093/nar/8.16.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Staden R. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 1982 Aug 11;10(15):4731–4751. doi: 10.1093/nar/10.15.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stanley E. R., Chen D. M., Lin H. S. Induction of macrophage production and proliferation by a purified colony stimulating factor. Nature. 1978 Jul 13;274(5667):168–170. doi: 10.1038/274168a0. [DOI] [PubMed] [Google Scholar]
  58. Taniguchi T., Matsui H., Fujita T., Takaoka C., Kashima N., Yoshimoto R., Hamuro J. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983 Mar 24;302(5906):305–310. doi: 10.1038/302305a0. [DOI] [PubMed] [Google Scholar]
  59. Thimmappaya B., Weinberger C., Schneider R. J., Shenk T. Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection. Cell. 1982 Dec;31(3 Pt 2):543–551. doi: 10.1016/0092-8674(82)90310-5. [DOI] [PubMed] [Google Scholar]
  60. Walter P., Blobel G. Signal recognition particle contains a 7S RNA essential for protein translocation across the endoplasmic reticulum. Nature. 1982 Oct 21;299(5885):691–698. doi: 10.1038/299691a0. [DOI] [PubMed] [Google Scholar]
  61. Welte K., Platzer E., Lu L., Gabrilove J. L., Levi E., Mertelsmann R., Moore M. A. Purification and biochemical characterization of human pluripotent hematopoietic colony-stimulating factor. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1526–1530. doi: 10.1073/pnas.82.5.1526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Wilbur W. J., Lipman D. J. Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci U S A. 1983 Feb;80(3):726–730. doi: 10.1073/pnas.80.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]
  64. Yokota T., Arai N., Lee F., Rennick D., Mosmann T., Arai K. Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: expression of T-cell growth-factor activity after transfection of monkey cells. Proc Natl Acad Sci U S A. 1985 Jan;82(1):68–72. doi: 10.1073/pnas.82.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Yokota T., Lee F., Rennick D., Hall C., Arai N., Mosmann T., Nabel G., Cantor H., Arai K. Isolation and characterization of a mouse cDNA clone that expresses mast-cell growth-factor activity in monkey cells. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1070–1074. doi: 10.1073/pnas.81.4.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]

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