Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Oct 1;168(4):1255–1270. doi: 10.1084/jem.168.4.1255

Identification of an alternative polyadenylation site in the human C3b/C4b receptor (complement receptor type 1) transcriptional unit and prediction of a secreted form of complement receptor type 1

PMCID: PMC2189081  PMID: 2971757

Abstract

The human C3b/C4b receptor or complement receptor type one (CR1) is an approximately 200-kD single chain membrane glycoprotein of human peripheral blood cells that mediates the binding, processing, and transport of C3b-bearing immune complexes and regulates the activity of the complement cascade. Analysis of partial cDNA clones has shown that the COOH terminus is composed predominantly of three tandemly repeated regions of 450 amino acids each (15). In this report, we present a cDNA sequence that encodes the NH2 terminus of CR1. It appears to have been derived from an alternatively processed transcript, caused by polyadenylation occurring at a site within an intron in the CR1 transcriptional unit. The resulting truncated messenger carries an open reading frame that would produce a short, secreted CR1 form. We present genomic sequences and Northern blots which support this hypothesis and we propose that the NH2-terminal end of CR1 is a likely location for active sites. In addition, we report evidence for a CR1-like sequence in the human genome and we present a model for the organization of CR1.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Alsenz J., Schulz T. F., Lambris J. D., Sim R. B., Dierich M. P. Structural and functional analysis of the complement component factor H with the use of different enzymes and monoclonal antibodies to factor H. Biochem J. 1985 Dec 15;232(3):841–850. doi: 10.1042/bj2320841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Atkinson J. P., Jones E. A. Biosynthesis of the human C3b/C4b receptor during differentiation of the HL-60 cell line. Identification and characterization of a precursor molecule. J Clin Invest. 1984 Nov;74(5):1649–1657. doi: 10.1172/JCI111581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumm J. W. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. doi: 10.1126/science.847462. [DOI] [PubMed] [Google Scholar]
  5. Caras I. W., Davitz M. A., Rhee L., Weddell G., Martin D. W., Jr, Nussenzweig V. Cloning of decay-accelerating factor suggests novel use of splicing to generate two proteins. Nature. 1987 Feb 5;325(6104):545–549. doi: 10.1038/325545a0. [DOI] [PubMed] [Google Scholar]
  6. Carroll M. C., Alicot E. M., Katzman P. J., Klickstein L. B., Smith J. A., Fearon D. T. Organization of the genes encoding complement receptors type 1 and 2, decay-accelerating factor, and C4-binding protein in the RCA locus on human chromosome 1. J Exp Med. 1988 Apr 1;167(4):1271–1280. doi: 10.1084/jem.167.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chung L. P., Reid K. B. Structural and functional studies on C4b-binding protein, a regulatory component of the human complement system. Biosci Rep. 1985 Oct-Nov;5(10-11):855–865. doi: 10.1007/BF01119897. [DOI] [PubMed] [Google Scholar]
  8. Dahlbäck B., Smith C. A., Müller-Eberhard H. J. Visualization of human C4b-binding protein and its complexes with vitamin K-dependent protein S and complement protein C4b. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3461–3465. doi: 10.1073/pnas.80.11.3461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Discipio R. G., Hugli T. E. Circular dichroism studies of human factor H. A regulatory component of the complement system. Biochim Biophys Acta. 1982 Dec 6;709(1):58–64. doi: 10.1016/0167-4838(82)90421-6. [DOI] [PubMed] [Google Scholar]
  10. Dykman T. R., Cole J. L., Iida K., Atkinson J. P. Polymorphism of human erythrocyte C3b/C4b receptor. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1698–1702. doi: 10.1073/pnas.80.6.1698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dykman T. R., Hatch J. A., Aqua M. S., Atkinson J. P. Polymorphism of the C3b/C4b receptor (CR1): characterization of a fourth allele. J Immunol. 1985 Mar;134(3):1787–1789. [PubMed] [Google Scholar]
  12. Dykman T. R., Hatch J. A., Atkinson J. P. Polymorphism of the human C3b/C4b receptor. Identification of a third allele and analysis of receptor phenotypes in families and patients with systemic lupus erythematosus. J Exp Med. 1984 Mar 1;159(3):691–703. doi: 10.1084/jem.159.3.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Early P., Rogers J., Davis M., Calame K., Bond M., Wall R., Hood L. Two mRNAs can be produced from a single immunoglobulin mu gene by alternative RNA processing pathways. Cell. 1980 Jun;20(2):313–319. doi: 10.1016/0092-8674(80)90617-0. [DOI] [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Gilmore-Hebert M., Wall R. Nuclear RNA precursors in the processing pathway to MOPC 21 kappa light chain messenger RNA. J Mol Biol. 1979 Dec 25;135(4):879–891. doi: 10.1016/0022-2836(79)90517-5. [DOI] [PubMed] [Google Scholar]
  16. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holers V. M., Chaplin D. D., Leykam J. F., Gruner B. A., Kumar V., Atkinson J. P. Human complement C3b/C4b receptor (CR1) mRNA polymorphism that correlates with the CR1 allelic molecular weight polymorphism. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2459–2463. doi: 10.1073/pnas.84.8.2459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holers V. M., Seya T., Brown E., O'Shea J. J., Atkinson J. P. Structural and functional studies on the human C3b/C4b receptor (CR1) purified by affinity chromatography using a monoclonal antibody. Complement. 1986;3(2):63–78. doi: 10.1159/000467882. [DOI] [PubMed] [Google Scholar]
  19. Hong K., Kinoshita T., Dohi Y., Inoue K. Effect of trypsinization on the activity of human factor H. J Immunol. 1982 Aug;129(2):647–652. [PubMed] [Google Scholar]
  20. Klickstein L. B., Wong W. W., Smith J. A., Weis J. H., Wilson J. G., Fearon D. T. Human C3b/C4b receptor (CR1). Demonstration of long homologous repeating domains that are composed of the short consensus repeats characteristics of C3/C4 binding proteins. J Exp Med. 1987 Apr 1;165(4):1095–1112. doi: 10.1084/jem.165.4.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kristensen T., D'Eustachio P., Ogata R. T., Chung L. P., Reid K. B., Tack B. F. The superfamily of C3b/C4b-binding proteins. Fed Proc. 1987 May 15;46(7):2463–2469. [PubMed] [Google Scholar]
  22. Kristensen T., Wetsel R. A., Tack B. F. Structural analysis of human complement protein H: homology with C4b binding protein, beta 2-glycoprotein I, and the Ba fragment of B2. J Immunol. 1986 May 1;136(9):3407–3411. [PubMed] [Google Scholar]
  23. Lai C. J., Dhar R., Khoury G. Mapping the spliced and unspliced late lytic SV40 RNAs. Cell. 1978 Aug;14(4):971–982. doi: 10.1016/0092-8674(78)90351-3. [DOI] [PubMed] [Google Scholar]
  24. Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
  25. Lozier J., Takahashi N., Putnam F. W. Complete amino acid sequence of human plasma beta 2-glycoprotein I. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3640–3644. doi: 10.1073/pnas.81.12.3640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lublin D. M., Griffith R. C., Atkinson J. P. Influence of glycosylation on allelic and cell-specific Mr variation, receptor processing, and ligand binding of the human complement C3b/C4b receptor. J Biol Chem. 1986 May 5;261(13):5736–5744. [PubMed] [Google Scholar]
  27. Lublin D. M., Lemons R. S., Le Beau M. M., Holers V. M., Tykocinski M. L., Medof M. E., Atkinson J. P. The gene encoding decay-accelerating factor (DAF) is located in the complement-regulatory locus on the long arm of chromosome 1. J Exp Med. 1987 Jun 1;165(6):1731–1736. doi: 10.1084/jem.165.6.1731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maeda N., Yang F., Barnett D. R., Bowman B. H., Smithies O. Duplication within the haptoglobin Hp2 gene. Nature. 1984 May 10;309(5964):131–135. doi: 10.1038/309131a0. [DOI] [PubMed] [Google Scholar]
  29. Mather E. L., Nelson K. J., Haimovich J., Perry R. P. Mode of regulation of immunoglobulin mu- and delta-chain expression varies during B-lymphocyte maturation. Cell. 1984 Feb;36(2):329–338. doi: 10.1016/0092-8674(84)90226-5. [DOI] [PubMed] [Google Scholar]
  30. Medof M. E., Lublin D. M., Holers V. M., Ayers D. J., Getty R. R., Leykam J. F., Atkinson J. P., Tykocinski M. L. Cloning and characterization of cDNAs encoding the complete sequence of decay-accelerating factor of human complement. Proc Natl Acad Sci U S A. 1987 Apr;84(7):2007–2011. doi: 10.1073/pnas.84.7.2007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nevins J. R., Darnell J. E., Jr Steps in the processing of Ad2 mRNA: poly(A)+ nuclear sequences are conserved and poly(A) addition precedes splicing. Cell. 1978 Dec;15(4):1477–1493. doi: 10.1016/0092-8674(78)90071-5. [DOI] [PubMed] [Google Scholar]
  32. Nevins J. R., Wilson M. C. Regulation of adenovirus-2 gene expression at the level of transcriptional termination and RNA processing. Nature. 1981 Mar 12;290(5802):113–118. doi: 10.1038/290113a0. [DOI] [PubMed] [Google Scholar]
  33. Perkins S. J., Chung L. P., Reid K. B. Unusual ultrastructure of complement-component-C4b-binding protein of human complement by synchrotron X-ray scattering and hydrodynamic analysis. Biochem J. 1986 Feb 1;233(3):799–807. doi: 10.1042/bj2330799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Podell D. N., Abraham G. N. A technique for the removal of pyroglutamic acid from the amino terminus of proteins using calf liver pyroglutamate amino peptidase. Biochem Biophys Res Commun. 1978 Mar 15;81(1):176–185. doi: 10.1016/0006-291x(78)91646-7. [DOI] [PubMed] [Google Scholar]
  35. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  36. Rey-Campos J., Rubinstein P., Rodriguez de Cordoba S. A physical map of the human regulator of complement activation gene cluster linking the complement genes CR1, CR2, DAF, and C4BP. J Exp Med. 1988 Feb 1;167(2):664–669. doi: 10.1084/jem.167.2.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ripoche J., Day A. J., Harris T. J., Sim R. B. The complete amino acid sequence of human complement factor H. Biochem J. 1988 Jan 15;249(2):593–602. doi: 10.1042/bj2490593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rodriguez de Cordoba S., Lublin D. M., Rubinstein P., Atkinson J. P. Human genes for three complement components that regulate the activation of C3 are tightly linked. J Exp Med. 1985 May 1;161(5):1189–1195. doi: 10.1084/jem.161.5.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rosenfeld M. G., Amara S. G., Evans R. M. Alternative RNA processing: determining neuronal phenotype. Science. 1984 Sep 21;225(4668):1315–1320. doi: 10.1126/science.6089345. [DOI] [PubMed] [Google Scholar]
  40. Ross G. D., Medof M. E. Membrane complement receptors specific for bound fragments of C3. Adv Immunol. 1985;37:217–267. doi: 10.1016/s0065-2776(08)60341-7. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Schibler U., Marcu K. B., Perry R. P. The synthesis and processing of the messenger RNAs specifying heavy and light chain immunoglobulins in MPC-11 cells. Cell. 1978 Dec;15(4):1495–1509. doi: 10.1016/0092-8674(78)90072-7. [DOI] [PubMed] [Google Scholar]
  43. Schulz T. F., Schwäble W., Stanley K. K., Weiss E., Dierich M. P. Human complement factor H: isolation of cDNA clones and partial cDNA sequence of the 38-kDa tryptic fragment containing the binding site for C3b. Eur J Immunol. 1986 Nov;16(11):1351–1355. doi: 10.1002/eji.1830161107. [DOI] [PubMed] [Google Scholar]
  44. Van Dyne S., Holers V. M., Lublin D. M., Atkinson J. P. The polymorphism of the C3b/C4b receptor in the normal population and in patients with systemic lupus erythematosus. Clin Exp Immunol. 1987 Jun;68(3):570–579. [PMC free article] [PubMed] [Google Scholar]
  45. Weis J. H., Morton C. C., Bruns G. A., Weis J. J., Klickstein L. B., Wong W. W., Fearon D. T. A complement receptor locus: genes encoding C3b/C4b receptor and C3d/Epstein-Barr virus receptor map to 1q32. J Immunol. 1987 Jan 1;138(1):312–315. [PubMed] [Google Scholar]
  46. Wong W. W., Jack R. M., Smith J. A., Kennedy C. A., Fearon D. T. Rapid purification of the human C3b/C4b receptor (CR1) by monoclonal antibody affinity chromatography. J Immunol Methods. 1985 Oct 10;82(2):303–313. doi: 10.1016/0022-1759(85)90362-x. [DOI] [PubMed] [Google Scholar]
  47. Wong W. W., Kennedy C. A., Bonaccio E. T., Wilson J. G., Klickstein L. B., Weis J. H., Fearon D. T. Analysis of multiple restriction fragment length polymorphisms of the gene for the human complement receptor type I. Duplication of genomic sequences occurs in association with a high molecular mass receptor allotype. J Exp Med. 1986 Nov 1;164(5):1531–1546. doi: 10.1084/jem.164.5.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wong W. W., Klickstein L. B., Smith J. A., Weis J. H., Fearon D. T. Identification of a partial cDNA clone for the human receptor for complement fragments C3b/C4b. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7711–7715. doi: 10.1073/pnas.82.22.7711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wong W. W., Wilson J. G., Fearon D. T. Genetic regulation of a structural polymorphism of human C3b receptor. J Clin Invest. 1983 Aug;72(2):685–693. doi: 10.1172/JCI111018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Yoon S. H., Fearon D. T. Characterization of a soluble form of the C3b/C4b receptor (CR1) in human plasma. J Immunol. 1985 May;134(5):3332–3338. [PubMed] [Google Scholar]
  51. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES