Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1982 Dec;2(12):1492–1500. doi: 10.1128/mcb.2.12.1492

Inhibition of Adenovirus DNA Synthesis In Vitro by Sera from Patients with Systemic Lupus Erythematosus

Marshall S Horwitz 1,2,3,*, Beth R Friefeld 1, Harold D Keiser 4
PMCID: PMC369958  PMID: 14582191

Abstract

Sera containing antinuclear antibodies from patients with systemic lupus erythematosus (SLE) and related disorders were tested for their effect on the synthesis of adenovirus (Ad) DNA in an in vitro replication system. After being heated at 60°C for 1 h, some sera from patients with SLE inhibited Ad DNA synthesis by 60 to 100%. Antibodies to double-stranded DNA were present in 15 of the 16 inhibitory sera, and inhibitory activity copurified with anti-double-stranded DNA in the immunoglobulin G fraction. These SLE sera did not inhibit the DNA polymerases α, β, γ and had no antibody to the 72,000-dalton DNA-binding protein necessary for Ad DNA synthesis. The presence of antibodies to single-stranded DNA and a variety of saline-extractable antigens (Sm, Ha, nRNP, and rRNP) did not correlate with SLE serum inhibitory activity. Methods previously developed for studying the individual steps in Ad DNA replication were used to determine the site of inhibition by the SLE sera that contained antibody to double-stranded DNA. Concentrations of the SLE inhibitor that decreased the elongation of Ad DNA by greater than 85% had no effect on either the initiation of Ad DNA synthesis or the polymerization of the first 26 deoxyribonucleotides.

Full text

PDF
1492

Images in this article

1495Image
on p.1495
1496Image
on p.1496
1496Image
on p.1496
1497Image
on p.1497

Selected References

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

  1. Alarcon-Segovia D., Ruiz-Arguelles A., Fishbein E. Antibody to nuclear ribonucleoprotein penetrates live human mononuclear cells through Fc receptors. Nature. 1978 Jan 5;271(5640):67–69. doi: 10.1038/271067a0. [DOI] [PubMed] [Google Scholar]
  2. Alarcón-Segovia D., Ruíz-Argüelles A., Fishbein E. Antibody penetration into living cells. I. Intranuclear immunoglobulin in peripheral blood mononuclear cells in mixed connective tissue disease and systemic lupus erythematosus. Clin Exp Immunol. 1979 Mar;35(3):364–375. [PMC free article] [PubMed] [Google Scholar]
  3. Arrand J. R., Roberts R. J. The nucleotide sequences at the termini of adenovirus-2 DNA. J Mol Biol. 1979 Mar 15;128(4):577–594. doi: 10.1016/0022-2836(79)90294-8. [DOI] [PubMed] [Google Scholar]
  4. CINADER B. Antibody to enzymes--a three-component system. Introduction: immunochemistry of enzymes. Ann N Y Acad Sci. 1963 May 8;103:495–548. doi: 10.1111/j.1749-6632.1963.tb53717.x. [DOI] [PubMed] [Google Scholar]
  5. Challberg M. D., Desiderio S. V., Kelly T. J., Jr Adenovirus DNA replication in vitro: characterization of a protein covalently linked to nascent DNA strands. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5105–5109. doi: 10.1073/pnas.77.9.5105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Challberg M. D., Kelly T. J., Jr Adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Feb;76(2):655–659. doi: 10.1073/pnas.76.2.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Challberg M. D., Ostrove J. M., Kelly T. J., Jr Initiation of adenovirus DNA replication: detection of covalent complexes between nucleotide and the 80-kilodalton terminal protein. J Virol. 1982 Jan;41(1):265–270. doi: 10.1128/jvi.41.1.265-270.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Enomoto T., Lichy J. H., Ikeda J. E., Hurwitz J. Adenovirus DNA replication in vitro: purification of the terminal protein in a functional form. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6779–6783. doi: 10.1073/pnas.78.11.6779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ginsberg B., Keiser H. A Millipore filter assay for antibodies to native DNA in sera of patients with systemic lupus erythematosus. Arthritis Rheum. 1973 Mar-Apr;16(2):199–207. doi: 10.1002/art.1780160210. [DOI] [PubMed] [Google Scholar]
  10. Horwitz M. S., Ariga H. Multiple rounds of adenovirus DNA synthesis in vitro. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1476–1480. doi: 10.1073/pnas.78.3.1476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horwitz M. S. Bidirectional replication of adenovirus type 2 DNA. J Virol. 1976 Apr;18(1):307–315. doi: 10.1128/jvi.18.1.307-315.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horwitz M. S., Scharff M. D., Maizel J. V., Jr Synthesis and assembly of adenovirus 2. I. Polypeptide synthesis, assembly of capsomeres, and morphogenesis of the virion. Virology. 1969 Dec;39(4):682–694. doi: 10.1016/0042-6822(69)90006-3. [DOI] [PubMed] [Google Scholar]
  13. Horwitz M. S. Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4291–4295. doi: 10.1073/pnas.75.9.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ikeda J. E., Enomoto T., Hurwitz J. Replication of adenovirus DNA-protein complex with purified proteins. Proc Natl Acad Sci U S A. 1981 Feb;78(2):884–888. doi: 10.1073/pnas.78.2.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kaplan L. M., Ariga H., Hurwitz J., Horwitz M. S. Complementation of the temperature-sensitive defect in H5ts125 adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5534–5538. doi: 10.1073/pnas.76.11.5534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Keiser H. D., Weinstein J. The detection and identification of antibodies to saline extractable nuclear antigens by counterimmunoelectrophoresis. Arthritis Rheum. 1980 Sep;23(9):1026–1035. doi: 10.1002/art.1780230910. [DOI] [PubMed] [Google Scholar]
  17. Keiser H. Preparation of 125I-labeled native DNA for use in radioimmunoassays for anti-native-DNA antibodies. Arthritis Rheum. 1973 Jul-Aug;16(4):468–470. doi: 10.1002/art.1780160406. [DOI] [PubMed] [Google Scholar]
  18. Kekwick R. A. The serum proteins in multiple myelomatosis. Biochem J. 1940 Sep;34(8-9):1248–1257. doi: 10.1042/bj0341248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kessler S. W. Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent. J Immunol. 1976 Nov;117(5 Pt 1):1482–1490. [PubMed] [Google Scholar]
  20. Klajman A., Farkash R., Myers B. D. Demonstration of antibodies to deoxyribonucleic acid by counterelectrophoresis. J Immunol. 1973 Oct;111(4):1136–1139. [PubMed] [Google Scholar]
  21. Koffler D., Carr R., Agnello V., Thoburn R., Kunkel H. G. Antibodies to polynucleotides in human sera: antigenic specificity and relation to disease. J Exp Med. 1971 Jul 1;134(1):294–312. doi: 10.1084/jem.134.1.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lechner R. L., Kelly T. J., Jr The structure of replicating adenovirus 2 DNA molecules. Cell. 1977 Dec;12(4):1007–1020. doi: 10.1016/0092-8674(77)90165-9. [DOI] [PubMed] [Google Scholar]
  23. Lerner M. R., Boyle J. A., Hardin J. A., Steitz J. A. Two novel classes of small ribonucleoproteins detected by antibodies associated with lupus erythematosus. Science. 1981 Jan 23;211(4480):400–402. doi: 10.1126/science.6164096. [DOI] [PubMed] [Google Scholar]
  24. Lichy J. H., Horwitz M. S., Hurwitz J. Formation of a covalent complex between the 80,000-dalton adenovirus terminal protein and 5'-dCMP in vitro. Proc Natl Acad Sci U S A. 1981 May;78(5):2678–2682. doi: 10.1073/pnas.78.5.2678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Longiaru M., Ikeda J. E., Jarkovsky Z., Horwitz S. B., Horwitz M. S. The effect of aphidicolin on adenovirus DNA synthesis. Nucleic Acids Res. 1979 Jul 25;6(10):3369–3386. doi: 10.1093/nar/6.10.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Maizel J. V., Jr, White D. O., Scharff M. D. The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology. 1968 Sep;36(1):115–125. doi: 10.1016/0042-6822(68)90121-9. [DOI] [PubMed] [Google Scholar]
  27. Nakamura R. M., Tan E. M. Recent progress in the study of autoantibodies to nuclear antigens. Hum Pathol. 1978 Jan;9(1):85–91. doi: 10.1016/s0046-8177(78)80010-0. [DOI] [PubMed] [Google Scholar]
  28. Ng C. W., Gregory K. F. Antibody to lactate dehydrogenase. II. Uptake, accumulation and specific intracellular action in malignant cells. Biochim Biophys Acta. 1968 Nov 12;170(1):45–53. doi: 10.1016/0304-4165(68)90159-1. [DOI] [PubMed] [Google Scholar]
  29. Papalian M., Lafer E., Wong R., Stollar B. D. Reaction of systemic lupus erythematosus antinative DNA antibodies with native DNA fragments from 20 to 1,200 base pairs. J Clin Invest. 1980 Feb;65(2):469–477. doi: 10.1172/JCI109690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Paronetto F., Koffler D. Immunofluorescent localization of immunoglobulins, complement, and fibrinogen in human diseases. I. Systemic lupus erythematosus. J Clin Invest. 1965 Oct;44(10):1657–1664. doi: 10.1172/JCI105272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rekosh D. M., Russell W. C., Bellet A. J., Robinson A. J. Identification of a protein linked to the ends of adenovirus DNA. Cell. 1977 Jun;11(2):283–295. doi: 10.1016/0092-8674(77)90045-9. [DOI] [PubMed] [Google Scholar]
  32. Robinson A. J., Younghusband H. B., Bellett A. J. A circula DNA-protein complex from adenoviruses. Virology. 1973 Nov;56(1):54–69. doi: 10.1016/0042-6822(73)90287-0. [DOI] [PubMed] [Google Scholar]
  33. Sharp G. C., Irvin W. S., LaRoque R. L., Velez C., Daly V., Kaiser A. D., Holman H. R. Association of autoantibodies to different nuclear antigens with clinical patterns of rheumatic disease and responsiveness to therapy. J Clin Invest. 1971 Feb;50(2):350–359. doi: 10.1172/JCI106502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sharp P. A., Moore C., Haverty J. L. The infectivity of adenovirus 5 DNA-protein complex. Virology. 1976 Dec;75(2):442–456. doi: 10.1016/0042-6822(76)90042-8. [DOI] [PubMed] [Google Scholar]
  35. Shinagawa M., Padmanabhan R. Nucleotide sequence at the inverted terminal repetition of adenovirus type 2 DNA. Biochem Biophys Res Commun. 1979 Apr 13;87(3):671–678. doi: 10.1016/0006-291x(79)92011-4. [DOI] [PubMed] [Google Scholar]
  36. Shu S., Provost T., Croxdale M. B., Reichlin M., Beutner E. H. Nuclear deposits of immunoglobulins in skin of patients with systemic lupus erythematosus. Clin Exp Immunol. 1977 Feb;27(2):238–244. [PMC free article] [PubMed] [Google Scholar]
  37. Svec K. H., Blair J. D., Kaplan M. H. Immunopathologic studies of systemic lupus erythematosus (SLE). I. Tissue-bound immunoglobulins in relation to serum antinuclear immunoglobulins in systemic lupus and in chronic liver disease with LE cell factor. J Clin Invest. 1967 Apr;46(4):558–568. doi: 10.1172/JCI105557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tan E. M., Kunkel H. G. An immunofluorescent study of the skin lesions in systemic lupus erythematosus. Arthritis Rheum. 1966 Feb;9(1):37–46. doi: 10.1002/art.1780090105. [DOI] [PubMed] [Google Scholar]
  39. Varesio L., Cappuccinelli P., Forni G. Inhibition of replication of virus-transformed fibroblasts by antibodies to RNA. Cancer Res. 1975 Dec;35(12):3558–3565. [PubMed] [Google Scholar]
  40. Wells J. V., Webb J., Van Deventer M., Fry B., Pollard K. M., Raftos J., Monk W., Nelson D. S. In vivo anti-nuclear antibodies in epithelial biopsies in SLE and other connective tissue diseases. Clin Exp Immunol. 1979 Dec;38(3):424–435. [PMC free article] [PubMed] [Google Scholar]
  41. Winnacker E. L. Adenovirus DNA: structure and function of a novel replicon. Cell. 1978 Aug;14(4):761–773. doi: 10.1016/0092-8674(78)90332-x. [DOI] [PubMed] [Google Scholar]
  42. Yang V. W., Lerner M. R., Steitz J. A., Flint S. J. A small nuclear ribonucleoprotein is required for splicing of adenoviral early RNA sequences. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1371–1375. doi: 10.1073/pnas.78.3.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES