Murine erythropoietin gene: cloning, expression, and human gene homology.
PDF (1.8M)
Journal: 1986/December - Molecular and Cellular Biology
ISSN: 0270-7306
PUBMED: 3773894
Abstract:
The gene for murine erythropoietin (EPO) was isolated from a mouse genomic library with a human EPO cDNA probe. Nucleotide sequence analysis permitted the identification of the murine EPO coding sequence and the prediction of the encoded amino acid sequence based on sequence conservation between the mouse and human EPO genes. Both the coding DNA and the amino acid sequences were 80% conserved between the two species. Transformation of COS-1 cells with a mammalian cell expression vector containing the murine EPO coding region resulted in secretion of murine EPO with biological activity on both murine and human erythroid progenitor cells. The transcription start site for the murine EPO gene in kidneys was determined. This permitted tentative identification of the transcription control region. The region included 140 base pairs upstream of the cap site which was over 90% conserved between the murine and human genes. Surprisingly, the first intron and much of the 5'- and 3'-untranslated sequences were also substantially conserved between the genes of the two species.
Open in
PUBMED | PMC | Google Scholar | Wikipedia
Relations:
Content
Citations
(31)
References
(40)
Chemicals
(1)
Genes
(1)
Organisms
(4)
Processes
(6)
Anatomy
(1)
Similar articles
Articles by the same authors
Discussion board
Mol Cell Biol 6(3): 849-858
PMID: 3773894

Murine erythropoietin gene: cloning, expression, and human gene homology.

Abstract

The gene for murine erythropoietin (EPO) was isolated from a mouse genomic library with a human EPO cDNA probe. Nucleotide sequence analysis permitted the identification of the murine EPO coding sequence and the prediction of the encoded amino acid sequence based on sequence conservation between the mouse and human EPO genes. Both the coding DNA and the amino acid sequences were 80% conserved between the two species. Transformation of COS-1 cells with a mammalian cell expression vector containing the murine EPO coding region resulted in secretion of murine EPO with biological activity on both murine and human erythroid progenitor cells. The transcription start site for the murine EPO gene in kidneys was determined. This permitted tentative identification of the transcription control region. The region included 140 base pairs upstream of the cap site which was over 90% conserved between the murine and human genes. Surprisingly, the first intron and much of the 5'- and 3'-untranslated sequences were also substantially conserved between the genes of the two species.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
icon of scanned page 849
849
icon of scanned page 850
850
icon of scanned page 851
851
icon of scanned page 852
852
icon of scanned page 853
853
icon of scanned page 854
854
icon of scanned page 855
855
icon of scanned page 856
856
icon of scanned page 857
857
icon of scanned page 858
858

Images in this article

Image
Image
on p.854
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Alexanian R, Vaughn WK, Ruchelman MW. Erythropoietin excretion in man following androgens. J Lab Clin Med. 1967 Nov;70(5):777–785. [PubMed] [Google Scholar]
  • Breathnach R, Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. [PubMed] [Google Scholar]
  • Cowan NJ, Dobner PR, Fuchs EV, Cleveland DW. Expression of human alpha-tubulin genes: interspecies conservation of 3' untranslated regions. Mol Cell Biol. 1983 Oct;3(10):1738–1745.[PMC free article] [PubMed] [Google Scholar]
  • Derman E, Krauter K, Walling L, Weinberger C, Ray M, Darnell JE., Jr Transcriptional control in the production of liver-specific mRNAs. Cell. 1981 Mar;23(3):731–739. [PubMed] [Google Scholar]
  • Dretzen G, Bellard M, Sassone-Corsi P, Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. [PubMed] [Google Scholar]
  • Dunn CD, Jarvis JH, Greenman JM. A quantitative bioassay for erythropoietin using mouse fetal liver cells. Exp Hematol. 1975 Jan;3(1):65–78. [PubMed] [Google Scholar]
  • Dynan WS, Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. Nature. 316(6031):774–778. [PubMed] [Google Scholar]
  • Eschbach JW, Mladenovic J, Garcia JF, Wahl PW, Adamson JW. The anemia of chronic renal failure in sheep. Response to erythropoietin-rich plasma in vivo. J Clin Invest. 1984 Aug;74(2):434–441.[PMC free article] [PubMed] [Google Scholar]
  • Favaloro J, Treisman R, Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. [PubMed] [Google Scholar]
  • Fung MC, Hapel AJ, Ymer S, Cohen DR, Johnson RM, Campbell HD, Young IG. Molecular cloning of cDNA for murine interleukin-3. Nature. 1984 Jan 19;307(5948):233–237. [PubMed] [Google Scholar]
  • Gillies SD, Morrison SL, Oi VT, Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. [PubMed] [Google Scholar]
  • Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. [PubMed] [Google Scholar]
  • GOLDWASSER E, JACOBSON LO, FRIED W, PLZAK LF. Studies on erythropoiesis. V. The effect of cobalt on the production of erythropoietin. Blood. 1958 Jan;13(1):55–60. [PubMed] [Google Scholar]
  • Gray PW, Goeddel DV. Cloning and expression of murine immune interferon cDNA. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5842–5846.[PMC free article] [PubMed] [Google Scholar]
  • Greenberger JS, Eckner RJ, Sakakeeny M, Marks P, Reid D, Nabel G, Hapel A, Ihle JN, Humphries KC. Interleukin 3-dependent hematopoietic progenitor cell lines. Fed Proc. 1983 Jul;42(10):2762–2771. [PubMed] [Google Scholar]
  • Hastie ND, Held WA, Toole JJ. Multiple genes coding for the androgen-regulated major urinary proteins of the mouse. Cell. 1979 Jun;17(2):449–457. [PubMed] [Google Scholar]
  • Higashi Y, Sokawa Y, Watanabe Y, Kawade Y, Ohno S, Takaoka C, Taniguchi T. Structure and expression of a cloned cDNA for mouse interferon-beta. J Biol Chem. 1983 Aug 10;258(15):9522–9529. [PubMed] [Google Scholar]
  • Jacobs K, Shoemaker C, Rudersdorf R, Neill SD, Kaufman RJ, Mufson A, Seehra J, Jones SS, Hewick R, Fritsch EF, et al. Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature. 313(6005):806–810. [PubMed] [Google Scholar]
  • JACOBSON LO, GOLDWASSER E, FRIED W, PLZAK L. Role of the kidney in erythropoiesis. Nature. 1957 Mar 23;179(4560):633–634. [PubMed] [Google Scholar]
  • 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. Nature. 313(6001):402–404. [PubMed] [Google Scholar]
  • Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872.[PMC free article] [PubMed] [Google Scholar]
  • KRANTZ SB, GALLIEN-LARTIGUE O, GOLDWASSER E. THE EFFECT OF ERYTHROPOIETIN UPON HEME SYNTHESIS BY MARROW CELLS IN VITRO. J Biol Chem. 1963 Dec;238:4085–4090. [PubMed] [Google Scholar]
  • Krystal G. A simple microassay for erythropoietin based on 3H-thymidine incorporation into spleen cells from phenylhydrazine treated mice. Exp Hematol. 1983 Aug;11(7):649–660. [PubMed] [Google Scholar]
  • Meijlink F, Curran T, Miller AD, Verma IM. Removal of a 67-base-pair sequence in the noncoding region of protooncogene fos converts it to a transforming gene. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4987–4991.[PMC free article] [PubMed] [Google Scholar]
  • Mellon P, Parker V, Gluzman Y, Maniatis T. Identification of DNA sequences required for transcription of the human alpha 1-globin gene in a new SV40 host-vector system. Cell. 1981 Dec;27(2 Pt 1):279–288. [PubMed] [Google Scholar]
  • Melton DW, Konecki DS, Brennand J, Caskey CT. Structure, expression, and mutation of the hypoxanthine phosphoribosyltransferase gene. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2147–2151.[PMC free article] [PubMed] [Google Scholar]
  • Murphy MJ, Jr, Miyake T. The role of glycoprotein hormones in the regulation of hematopoiesis. Nihon Ketsueki Gakkai Zasshi. 1983 Dec;46(7):1380–1396. [PubMed] [Google Scholar]
  • Naughton BA, Kaplan SM, Roy M, Burdowski AJ, Gordon AS, Piliero SJ. Hepatic regeneration and erythropoietin production in the rat. Science. 1977 Apr 15;196(4287):301–302. [PubMed] [Google Scholar]
  • Reynolds GA, Basu SK, Osborne TF, Chin DJ, Gil G, Brown MS, Goldstein JL, Luskey KL. HMG CoA reductase: a negatively regulated gene with unusual promoter and 5' untranslated regions. Cell. 1984 Aug;38(1):275–285. [PubMed] [Google Scholar]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467.[PMC free article] [PubMed] [Google Scholar]
  • Schooley JC, Mahlmann LJ. Stimulation of erythropoiesis in plethoric mice by prostaglandins and its inhibition by antierythropoietin. Proc Soc Exp Biol Med. 1971 Nov;138(2):523–524. [PubMed] [Google Scholar]
  • Shaw GD, Boll W, Taira H, Mantei N, Lengyel P, Weissmann C. Structure and expression of cloned murine IFN-alpha genes. Nucleic Acids Res. 1983 Feb 11;11(3):555–573.[PMC free article] [PubMed] [Google Scholar]
  • Sherwood JB, Goldwasser E. Extraction of erythropoietin from normal kidneys. Endocrinology. 1978 Sep;103(3):866–870. [PubMed] [Google Scholar]
  • Singer-Sam J, Keith DH, Tani K, Simmer RL, Shively L, Lindsay S, Yoshida A, Riggs AD. Sequence of the promoter region of the gene for human X-linked 3-phosphoglycerate kinase. Gene. 1984 Dec;32(3):409–417. [PubMed] [Google Scholar]
  • Sompayrac LM, Danna KJ. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578.[PMC free article] [PubMed] [Google Scholar]
  • Sytkowski AJ. Denaturation and renaturation of human erythropoietin. Biochem Biophys Res Commun. 1980 Sep 16;96(1):143–149. [PubMed] [Google Scholar]
  • Valerio D, Duyvesteyn MG, Dekker BM, Weeda G, Berkvens TM, van der Voorn L, van Ormondt H, van der Eb AJ. Adenosine deaminase: characterization and expression of a gene with a remarkable promoter. EMBO J. 1985 Feb;4(2):437–443.[PMC free article] [PubMed] [Google Scholar]
  • Wang FF, Kung CK, Goldwasser E. Some chemical properties of human erythropoietin. Endocrinology. 1985 Jun;116(6):2286–2292. [PubMed] [Google Scholar]
  • Wong GG, Witek JS, Temple PA, Wilkens KM, Leary AC, Luxenberg DP, Jones SS, Brown EL, Kay RM, Orr EC, et al. Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins. Science. 1985 May 17;228(4701):810–815. [PubMed] [Google Scholar]
  • Zanjani ED, Peterson EN, Gordon AS, Wasserman LR. Erythropoietin production in the fetus: role of the kidney and maternal anemia. J Lab Clin Med. 1974 Feb;83(2):281–287. [PubMed] [Google Scholar]
Copyright notice
Abstract
The gene for murine erythropoietin (EPO) was isolated from a mouse genomic library with a human EPO cDNA probe. Nucleotide sequence analysis permitted the identification of the murine EPO coding sequence and the prediction of the encoded amino acid sequence based on sequence conservation between the mouse and human EPO genes. Both the coding DNA and the amino acid sequences were 80% conserved between the two species. Transformation of COS-1 cells with a mammalian cell expression vector containing the murine EPO coding region resulted in secretion of murine EPO with biological activity on both murine and human erythroid progenitor cells. The transcription start site for the murine EPO gene in kidneys was determined. This permitted tentative identification of the transcription control region. The region included 140 base pairs upstream of the cap site which was over 90% conserved between the murine and human genes. Surprisingly, the first intron and much of the 5'- and 3'-untranslated sequences were also substantially conserved between the genes of the two species.
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.