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. 1996 Mar;70(3):1836–1844. doi: 10.1128/jvi.70.3.1836-1844.1996

Analysis of 15 adenovirus hexon proteins reveals the location and structure of seven hypervariable regions containing serotype-specific residues.

L Crawford-Miksza 1, D P Schnurr 1
PMCID: PMC190011  PMID: 8627708

Abstract

The first full-length hexon protein DNA and deduced amino acid sequences of a subgenus D adenovirus (AV) were determined from candidate AV48 (85-0844). Comprehensive comparison of this sequence with hexon protein sequences from human subgenera A, B, C, D, F, bovine AV3, and mouse AV1 revealed seven discrete hypervariable regions (HVRs) among the 250 variable residues in loops 1 and 2. These regions differed in length between serotypes, from 2 to 38 residues, and contained > 00% of hexon serotype-specific residues among human serotypes. Alignment with the published crystal structure of AV2 established the location and structure of the type-specific regions. Five HVRs were shown to be part of linear loops on the exposed surfaces of the protein, analogous to the serotype-specific loops or "puffs" in picornavirus capsid proteins. The HVRs were supported by a common framework of conserved residues, of which 68 to 75% were hydrophobic. Unique sequences were limited to the seven HVRs, so that one or more of these regions contain the type-specific neutralization epitopes. A neutralizing AV48 hexon-specific antiserum recognized linear peptides that corresponded to six HVRs by enzyme immunoassay. Affinity-purification removal of all peptide-reactive antibodies did not significantly decrease the neutralization titer. Eluted peptide-reactive antibodies did not neutralize. Human antisera that neutralized AV48 did not recognize linear peptides. Purified trimeric native hexon inhibited neutralization, but monomeric heat-denatured hexon did not. We conclude that the AV48 neutralization epitope(s) is complex and conformational.

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Selected References

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  1. Athappilly F. K., Murali R., Rux J. J., Cai Z., Burnett R. M. The refined crystal structure of hexon, the major coat protein of adenovirus type 2, at 2.9 A resolution. J Mol Biol. 1994 Sep 30;242(4):430–455. doi: 10.1006/jmbi.1994.1593. [DOI] [PubMed] [Google Scholar]
  2. Boulanger P. A., Puvion F. Large-scale preparation of soluble adenovirus hexon, penton and fiber antigens in highly purified form. Eur J Biochem. 1973 Nov 1;39(1):37–42. doi: 10.1111/j.1432-1033.1973.tb03100.x. [DOI] [PubMed] [Google Scholar]
  3. Crawford-Miksza L. K., Schnurr D. P. Quantitative colorimetric microneutralization assay for characterization of adenoviruses. J Clin Microbiol. 1994 Sep;32(9):2331–2334. doi: 10.1128/jcm.32.9.2331-2334.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davies D. R., Sheriff S., Padlan E. A. Antibody-antigen complexes. J Biol Chem. 1988 Aug 5;263(22):10541–10544. [PubMed] [Google Scholar]
  5. Hierholzer J. C. Adenoviruses in the immunocompromised host. Clin Microbiol Rev. 1992 Jul;5(3):262–274. doi: 10.1128/cmr.5.3.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hierholzer J. C., Adrian T., Anderson L. J., Wigand R., Gold J. W. Analysis of antigenically intermediate strains of subgenus B and D adenoviruses from AIDS patients. Arch Virol. 1988;103(1-2):99–115. doi: 10.1007/BF01319812. [DOI] [PubMed] [Google Scholar]
  7. Hierholzer J. C., Stone Y. O., Broderson J. R. Antigenic relationships among the 47 human adenoviruses determined in reference horse antisera. Arch Virol. 1991;121(1-4):179–197. doi: 10.1007/BF01316753. [DOI] [PubMed] [Google Scholar]
  8. Hierholzer J. C., Wigand R., Anderson L. J., Adrian T., Gold J. W. Adenoviruses from patients with AIDS: a plethora of serotypes and a description of five new serotypes of subgenus D (types 43-47). J Infect Dis. 1988 Oct;158(4):804–813. doi: 10.1093/infdis/158.4.804. [DOI] [PubMed] [Google Scholar]
  9. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  10. Horsfall A. C., Hay F. C., Soltys A. J., Jones M. G. Epitope mapping. Immunol Today. 1991 Jul;12(7):211–213. doi: 10.1016/0167-5699(91)90029-S. [DOI] [PubMed] [Google Scholar]
  11. Hu S. L., Hays W. W., Potts D. E. Sequence homology between bovine and human adenoviruses. J Virol. 1984 Feb;49(2):604–608. doi: 10.1128/jvi.49.2.604-608.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. JAWETZ E., THYGESON P., HANNA L., NICHOLAS A., KIMURA S. J. The etiology of epidemic keratoconjunctivitis. Am J Ophthalmol. 1957 Apr;43(4 Pt 2):79–83. doi: 10.1016/0002-9394(57)91483-6. [DOI] [PubMed] [Google Scholar]
  13. Kinloch R., Mackay N., Mautner V. Adenovirus hexon. Sequence comparison of subgroup C serotypes 2 and 5. J Biol Chem. 1984 May 25;259(10):6431–6436. [PubMed] [Google Scholar]
  14. Norrby E. The structural and functional diversity of Adenovirus capsid components. J Gen Virol. 1969 Sep;5(2):221–236. doi: 10.1099/0022-1317-5-2-221. [DOI] [PubMed] [Google Scholar]
  15. Novotný J., Handschumacher M., Haber E., Bruccoleri R. E., Carlson W. B., Fanning D. W., Smith J. A., Rose G. D. Antigenic determinants in proteins coincide with surface regions accessible to large probes (antibody domains). Proc Natl Acad Sci U S A. 1986 Jan;83(2):226–230. doi: 10.1073/pnas.83.2.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nuss J. M., Air G. M. Defining the requirements for an antibody epitope on influenza virus neuraminidase: how tolerant are protein epitopes? J Mol Biol. 1994 Jan 14;235(2):747–759. doi: 10.1006/jmbi.1994.1025. [DOI] [PubMed] [Google Scholar]
  17. Pring-Akerblom P., Adrian T. The hexon genes of adenoviruses of subgenus C: comparison of the variable regions. Res Virol. 1993 Mar-Apr;144(2):117–127. doi: 10.1016/s0923-2516(06)80020-8. [DOI] [PubMed] [Google Scholar]
  18. Pring-Akerblom P., Adrian T. Type- and group-specific polymerase chain reaction for adenovirus detection. Res Virol. 1994 Jan-Feb;145(1):25–35. doi: 10.1016/s0923-2516(07)80004-5. [DOI] [PubMed] [Google Scholar]
  19. Roberts M. M., White J. L., Grütter M. G., Burnett R. M. Three-dimensional structure of the adenovirus major coat protein hexon. Science. 1986 May 30;232(4754):1148–1151. doi: 10.1126/science.3704642. [DOI] [PubMed] [Google Scholar]
  20. Rossmann M. G., Arnold E., Erickson J. W., Frankenberger E. A., Griffith J. P., Hecht H. J., Johnson J. E., Kamer G., Luo M., Mosser A. G. Structure of a human common cold virus and functional relationship to other picornaviruses. Nature. 1985 Sep 12;317(6033):145–153. doi: 10.1038/317145a0. [DOI] [PubMed] [Google Scholar]
  21. Schnurr D., Dondero M. E. Two new candidate adenovirus serotypes. Intervirology. 1993;36(2):79–83. doi: 10.1159/000150325. [DOI] [PubMed] [Google Scholar]
  22. Seligman S. J. Serial deletion mapping by competition ELISA assay: characterization of a linear epitope in the V3 loop of HIV-1. AIDS Res Hum Retroviruses. 1994 Feb;10(2):149–156. doi: 10.1089/aid.1994.10.149. [DOI] [PubMed] [Google Scholar]
  23. Sprengel J., Schmitz B., Heuss-Neitzel D., Zock C., Doerfler W. Nucleotide sequence of human adenovirus type 12 DNA: comparative functional analysis. J Virol. 1994 Jan;68(1):379–389. doi: 10.1128/jvi.68.1.379-389.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Toogood C. I., Crompton J., Hay R. T. Antipeptide antisera define neutralizing epitopes on the adenovirus hexon. J Gen Virol. 1992 Jun;73(Pt 6):1429–1435. doi: 10.1099/0022-1317-73-6-1429. [DOI] [PubMed] [Google Scholar]
  25. Toogood C. I., Hay R. T. DNA sequence of the adenovirus type 41 hexon gene and predicted structure of the protein. J Gen Virol. 1988 Sep;69(Pt 9):2291–2301. doi: 10.1099/0022-1317-69-9-2291. [DOI] [PubMed] [Google Scholar]
  26. Toogood C. I., Murali R., Burnett R. M., Hay R. T. The adenovirus type 40 hexon: sequence, predicted structure and relationship to other adenovirus hexons. J Gen Virol. 1989 Dec;70(Pt 12):3203–3214. doi: 10.1099/0022-1317-70-12-3203. [DOI] [PubMed] [Google Scholar]
  27. Varga M. J., Weibull C., Everitt E. Infectious entry pathway of adenovirus type 2. J Virol. 1991 Nov;65(11):6061–6070. doi: 10.1128/jvi.65.11.6061-6070.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vogt R. F., Jr, Phillips D. L., Henderson L. O., Whitfield W., Spierto F. W. Quantitative differences among various proteins as blocking agents for ELISA microtiter plates. J Immunol Methods. 1987 Jul 16;101(1):43–50. doi: 10.1016/0022-1759(87)90214-6. [DOI] [PubMed] [Google Scholar]
  29. Weber J. M., Cai F., Murali R., Burnett R. M. Sequence and structural analysis of murine adenovirus type 1 hexon. J Gen Virol. 1994 Jan;75(Pt 1):141–147. doi: 10.1099/0022-1317-75-1-141. [DOI] [PubMed] [Google Scholar]
  30. Weiner A. J., Geysen H. M., Christopherson C., Hall J. E., Mason T. J., Saracco G., Bonino F., Crawford K., Marion C. D., Crawford K. A. Evidence for immune selection of hepatitis C virus (HCV) putative envelope glycoprotein variants: potential role in chronic HCV infections. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3468–3472. doi: 10.1073/pnas.89.8.3468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wigand R., Adrian T. Intermediate adenovirus strains of subgenus D occur in extensive variety. Med Microbiol Immunol. 1989;178(1):37–44. doi: 10.1007/BF00202290. [DOI] [PubMed] [Google Scholar]
  32. Wigand R., Bartha A., Dreizin R. S., Esche H., Ginsberg H. S., Green M., Hierholzer J. C., Kalter S. S., McFerran J. B., Pettersson U. Adenoviridae: second report. Intervirology. 1982;18(4):169–176. doi: 10.1159/000149322. [DOI] [PubMed] [Google Scholar]
  33. Wohlfart C. Neutralization of adenoviruses: kinetics, stoichiometry, and mechanisms. J Virol. 1988 Jul;62(7):2321–2328. doi: 10.1128/jvi.62.7.2321-2328.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van Oostrum J., Smith P. R., Mohraz M., Burnett R. M. The structure of the adenovirus capsid. III. Hexon packing determined from electron micrographs of capsid fragments. J Mol Biol. 1987 Nov 5;198(1):73–89. doi: 10.1016/0022-2836(87)90459-1. [DOI] [PubMed] [Google Scholar]

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