Human Genetics

, Volume 91, Issue 2, pp 157–162

Isolation of a new cDNA clone encoding an Rh polypeptide associated with the Rh blood group system

  • Eiji Kajii
  • Fuminori Umenishi
  • Sadahiko Iwamoto
  • Shigenori Ikemoto
Original Investigations

Abstract

The polymerase chain reaction (PCR) was used to amplify Rh-related cDNAs from erythroid cells cultured by the selective two-phase liquid culture system for human erythroid progenitors in peripheral blood. Direct sequencing based on PCR presents heterozygous bands. Two Rh polypeptide cDNAs have been isolated from the PCR products and tentatively designated RhPI cDNA and RhPII cDNA. Both cDNA clones have an open reading frame composed of 1251 nucleotides. The RhPI cDNA clone shows a single nucleotide substitution with no amino acid substitution compared with the published sequence. The RhPII cDNA clone, on the other hand, differs from the above by 41 nucleotide substitutions with the open reading frame, resulting in 31 amino acid substitutions.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Agre P, Cartron J-P (1991) Molecular biology of the Rh antigens. Blood 78:551–563Google Scholar
  2. Avent ND, Ridgwell K, Mawby WJ, Tanner MJA, Anstee DJ, Kumpel B (1988) Protein-sequence studies of Rh-related polypeptides suggest the presence of at least two groups of proteins which associate in the human red-cell membrane. Biochem J 256:1043–1046Google Scholar
  3. Avent N, Ridgwell K, Tanner MJA, Anstee DJ (1990) cDNA cloning of a 30kDa erythrocyte membrane protein associated with Rh (Rhesus)-blood-group-antigen expression. Biochem J 271:821–825Google Scholar
  4. Blanchard D, Bloy C, Hermand P, Cartron J-P, Saboori AM, Smith BL, Agre P (1988) Two-dimensional iodopeptide mapping demonstrates erythrocyte RhD, c, and E polypeptides are structurally homologous but nonidentical. Blood 72:1424–1427Google Scholar
  5. Bloy C, Blanchard D, Lambin P, Goossens D, Rouger P, Salmon C, Cartron J-P (1987) Human monoclonal antibody against Rh(D) antigens: partial characterization of the Rh(D) polypeptide from human erythrocytes. Blood 69:1491–1497Google Scholar
  6. Bloy C, Blanchard D, Dahr W, Beyreuther K, Salmon C, Cartron J-P (1988) Determination of the N-terminal sequence of human red cell Rh(D) polypeptide and demonstration that the Rh(D), (c), and (E) antigens are carried by distinct polypeptide chains. Blood 72:661–666Google Scholar
  7. Bruns GAP, Sherman SL (1989) Report of the committee on the genetic constitution of chromosome I. (10th International Workshop on Human Gene Mapping) Cytogenet Cell Genet 51:67–90Google Scholar
  8. Chérif-Zahar B, Bloy C, Le Van Kim C, Blanchard D, Bailly P, Hermand P, Salmon C, Cartron J-P, Colin Y (1990) Molecular cloning and protein structure of a human blood group Rh polypeptide. Proc Natl Acad Sci USA 87:6243–6247Google Scholar
  9. Chérif-Zahar B, Mattel MG, Le Van Kim C, Bailly P, Cartron J-P, Colin Y (1991) Localization of the human Rh blood group gene structure to chromosome region 1p34.3–1p36.1 by in situ hybridization. Hum Genet 86:398–400Google Scholar
  10. Colin Y, Chérif-Zahar B, Le Van Kim C, Raynal V, Van Huffel V, Cartron J-P (1991) Genetic basis of the RhD-positive and RhD-negative blood group polymorphism as determined by Southern analysis. Blood 78:2747–2752Google Scholar
  11. Erlich HA (1989) PCR technology: principles and application of DNA amplification. Stockton, New YorkGoogle Scholar
  12. Gahmberg CG (1982) Molecular identification of the human Rh0(D) antigen. FEBS Lett 140:93–97PubMedGoogle Scholar
  13. Green FA (1967) Erythrocyte membrane sulfhydryl groups and Rh antigen activity. Immunochemistry 4:247–257Google Scholar
  14. Green FA (1983) The mode of attenuation of erythrocyte membrane Rh0(D) antigen activity by 5. 5′-dithiobis-(2-nitrobenzoic acid) and protection against loss of activity by bound anti-Rh0(D) antibody. Mol Immunol 20:769–775Google Scholar
  15. Gyllensten UB, Erlich HA (1988) Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci USA 85:7652–7656Google Scholar
  16. Hartmann E, Rapoport TA, Lodish HF (1989) Predicting the orientation of eukaryotic membrane-spanning proteins. Proc Natl Acad Sci USA 86:5786–5790Google Scholar
  17. Hashimoto K, Nakaki Y, Amemiya Y (1986) Quantitation of antigens and immunoglobulins on red blood cells by flow cytometry. Immunohaematology 8:457–461Google Scholar
  18. Kajii E, Ikemoto S (1992) A new method for isolation of Rh polypeptide mRNAs using a liquid culture system. Proc Jpn Acad 68:51–56Google Scholar
  19. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132PubMedGoogle Scholar
  20. Leff SE, Rosenfeld MG, Evans RM (1986) Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem 55:1091–1096CrossRefPubMedGoogle Scholar
  21. Mollison PL, Engelfriet CP, Contreras M (1987) Blood transfusion in clinical medicine, 8th edn. Blackwell, OxfordGoogle Scholar
  22. Moore S, Woodrow CF, McClelland DBL (1982) Isolation of membrane components associated with human red cell antigens Rh(D), (c), (E) and Fya. Nature 295:529–531Google Scholar
  23. Petz LD, Garratty G (1980) Acquired immune hemolytic anemia. Churchill Livingstone, New YorkGoogle Scholar
  24. Saboori AM, Smith BL, Agre P (1988) Polymorphism in the Mr 32000 Rh protein purified from Rh(D) positive and negative erythrocytes. Proc Natl Acad Sci USA 85:4042–4045Google Scholar
  25. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NYGoogle Scholar
  26. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467PubMedGoogle Scholar
  27. Suda T, Suda J, Miura Y, Hayashi Y, Eguchi M, Tadokoro K, Saito M (1985) Clonal analysis of basophil differentiation in bone marrow cultures from Down's syndrome patient with megakaryoblastic leukemia. Blood 66:1278–1283Google Scholar
  28. Suyama K, Goldstein J, Aebersold R, Kent S (1991) Regarding the size of Rh proteins. Blood 77:411Google Scholar
  29. Tebbi K, Rubin S, Cowan DH, McCulloch EA (1976) A comparison of granulopoiesis in culture from blood and marrow cells of nonleukemic individuals and patients with acute leukemia. Blood 48:235–243Google Scholar
  30. Wada H, Suda T, Miura Y, Kajii E, Ikemoto S, Yawata Y (1990) Expression of major blood group antigens on human erythroid cells in a two phase liquid culture system. Blood 75:505–511Google Scholar
  31. Wang AM, Doyle MV, Mark DF (1989) Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci USA 86:9717–9721Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Eiji Kajii
    • 1
  • Fuminori Umenishi
    • 1
  • Sadahiko Iwamoto
    • 1
  • Shigenori Ikemoto
    • 1
  1. 1.Department of Legal Medicine and Human GeneticsJichi Medical SchoolTochigiJapan

Personalised recommendations