Abstract
The adenovirus fiber protein is responsible for attachment of the virion to cell surface receptors. The identity of the cellular receptor which mediates binding is unknown, although there is evidence suggesting that two distinct adenovirus receptors interact with the group C (adenovirus type 5 [Ad5]) and the group B (Ad3) adenoviruses. In order to define the determinants of adenovirus receptor specificity, we have carried out a series of competition binding experiments using recombinant native fiber polypeptides from Ad5 and Ad3 and chimeric fiber proteins in which the head domains of Ad5 and Ad3 were exchanged. Specific binding of fiber to HeLa cell receptors was assessed with radiolabeled protein synthesized in vitro, and by competition analysis with baculovirus-expressed fiber protein. Fiber produced in vitro was found as both monomer and trimer, but only the assembled trimers had receptor binding activity. Competition data support the conclusion that Ad5 and Ad3 interact with different cellular receptors. The Ad5 receptor distribution on several cell lines was assessed with a fiber binding flow cytometric assay. HeLa cells were found to express high levels of receptor, while CHO and human diploid fibroblasts did not. A chimeric fiber containing the Ad5 fiber head domain blocked the binding of Ad5 fiber but not Ad3 fiber. Similarly, a chimeric fiber containing the Ad3 fiber head blocked the binding of labeled Ad3 fiber but not Ad5 fiber. In addition, the isolated Ad3 fiber head domain competed effectively with labeled Ad3 fiber for binding to HeLa cell receptors. These results demonstrate that the determinants of receptor binding are located in the head domain of the fiber and that the isolated head domain is capable of trimerization and binding to cellular receptors. Our results also show that it is possible to change the receptor specificity of the fiber protein by manipulation of sequences contained in the head domain. Modification or replacement of the fiber head domain with novel ligands may permit adenovirus vectors with new receptor specificities which could be useful for targeted gene delivery in vivo to be engineered.
Full Text
The Full Text of this article is available as a PDF (322.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bai M., Campisi L., Freimuth P. Vitronectin receptor antibodies inhibit infection of HeLa and A549 cells by adenovirus type 12 but not by adenovirus type 2. J Virol. 1994 Sep;68(9):5925–5932. doi: 10.1128/jvi.68.9.5925-5932.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bai M., Harfe B., Freimuth P. Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells. J Virol. 1993 Sep;67(9):5198–5205. doi: 10.1128/jvi.67.9.5198-5205.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chroboczek J., Jacrot B. The sequence of adenovirus fiber: similarities and differences between serotypes 2 and 5. Virology. 1987 Dec;161(2):549–554. doi: 10.1016/0042-6822(87)90150-4. [DOI] [PubMed] [Google Scholar]
- Chu T. H., Martinez I., Sheay W. C., Dornburg R. Cell targeting with retroviral vector particles containing antibody-envelope fusion proteins. Gene Ther. 1994 Sep;1(5):292–299. [PubMed] [Google Scholar]
- Defer C., Belin M. T., Caillet-Boudin M. L., Boulanger P. Human adenovirus-host cell interactions: comparative study with members of subgroups B and C. J Virol. 1990 Aug;64(8):3661–3673. doi: 10.1128/jvi.64.8.3661-3673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devaux C., Adrian M., Berthet-Colominas C., Cusack S., Jacrot B. Structure of adenovirus fibre. I. Analysis of crystals of fibre from adenovirus serotypes 2 and 5 by electron microscopy and X-ray crystallography. J Mol Biol. 1990 Oct 20;215(4):567–588. doi: 10.1016/S0022-2836(05)80169-X. [DOI] [PubMed] [Google Scholar]
- Green N. M., Wrigley N. G., Russell W. C., Martin S. R., McLachlan A. D. Evidence for a repeating cross-beta sheet structure in the adenovirus fibre. EMBO J. 1983;2(8):1357–1365. doi: 10.1002/j.1460-2075.1983.tb01592.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henry L. J., Xia D., Wilke M. E., Deisenhofer J., Gerard R. D. Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli. J Virol. 1994 Aug;68(8):5239–5246. doi: 10.1128/jvi.68.8.5239-5246.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong J. S., Engler J. A. The amino terminus of the adenovirus fiber protein encodes the nuclear localization signal. Virology. 1991 Dec;185(2):758–767. doi: 10.1016/0042-6822(91)90547-o. [DOI] [PubMed] [Google Scholar]
- Horton R. M., Cai Z. L., Ho S. N., Pease L. R. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. Biotechniques. 1990 May;8(5):528–535. [PubMed] [Google Scholar]
- Kasahara N., Dozy A. M., Kan Y. W. Tissue-specific targeting of retroviral vectors through ligand-receptor interactions. Science. 1994 Nov 25;266(5189):1373–1376. doi: 10.1126/science.7973726. [DOI] [PubMed] [Google Scholar]
- Kidd A. H., Chroboczek J., Cusack S., Ruigrok R. W. Adenovirus type 40 virions contain two distinct fibers. Virology. 1993 Jan;192(1):73–84. doi: 10.1006/viro.1993.1009. [DOI] [PubMed] [Google Scholar]
- Kidd A. H., Erasmus M. J. Sequence characterization of the adenovirus 40 fiber gene. Virology. 1989 Sep;172(1):134–144. doi: 10.1016/0042-6822(89)90115-3. [DOI] [PubMed] [Google Scholar]
- Kidd A. H., Erasmus M. J., Tiemessen C. T. Fiber sequence heterogeneity in subgroup F adenoviruses. Virology. 1990 Nov;179(1):139–150. doi: 10.1016/0042-6822(90)90283-w. [DOI] [PubMed] [Google Scholar]
- Leone G., Maybaum L., Lee P. W. The reovirus cell attachment protein possesses two independently active trimerization domains: basis of dominant negative effects. Cell. 1992 Oct 30;71(3):479–488. doi: 10.1016/0092-8674(92)90516-f. [DOI] [PubMed] [Google Scholar]
- Lonberg-Holm K., Crowell R. L., Philipson L. Unrelated animal viruses share receptors. Nature. 1976 Feb 26;259(5545):679–681. doi: 10.1038/259679a0. [DOI] [PubMed] [Google Scholar]
- Louis N., Fender P., Barge A., Kitts P., Chroboczek J. Cell-binding domain of adenovirus serotype 2 fiber. J Virol. 1994 Jun;68(6):4104–4106. doi: 10.1128/jvi.68.6.4104-4106.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mathias P., Wickham T., Moore M., Nemerow G. Multiple adenovirus serotypes use alpha v integrins for infection. J Virol. 1994 Oct;68(10):6811–6814. doi: 10.1128/jvi.68.10.6811-6814.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Novelli A., Boulanger P. A. Assembly of adenovirus type 2 fiber synthesized in cell-free translation system. J Biol Chem. 1991 May 15;266(14):9299–9303. [PubMed] [Google Scholar]
- Novelli A., Boulanger P. A. Deletion analysis of functional domains in baculovirus-expressed adenovirus type 2 fiber. Virology. 1991 Nov;185(1):365–376. doi: 10.1016/0042-6822(91)90784-9. [DOI] [PubMed] [Google Scholar]
- Philipson L., Lonberg-Holm K., Pettersson U. Virus-receptor interaction in an adenovirus system. J Virol. 1968 Oct;2(10):1064–1075. doi: 10.1128/jvi.2.10.1064-1075.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pieniazek N. J., Slemenda S. B., Pieniazek D., Velarde J., Jr, Luftig R. B. Human enteric adenovirus type 41 (Tak) contains a second fiber protein gene. Nucleic Acids Res. 1990 Apr 11;18(7):1901–1901. doi: 10.1093/nar/18.7.1901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pieniazek N. J., Slemenda S. B., Pieniazek D., Velarde J., Jr, Luftig R. B. Sequence of human enteric adenovirus type 41 Tak fiber protein gene. Nucleic Acids Res. 1989 Nov 25;17(22):9474–9474. doi: 10.1093/nar/17.22.9474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Signäs C., Akusjärvi G., Pettersson U. Adenovirus 3 fiber polypeptide gene: implications for the structure of the fiber protein. J Virol. 1985 Feb;53(2):672–678. doi: 10.1128/jvi.53.2.672-678.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Stewart P. L., Burnett R. M., Cyrklaff M., Fuller S. D. Image reconstruction reveals the complex molecular organization of adenovirus. Cell. 1991 Oct 4;67(1):145–154. doi: 10.1016/0092-8674(91)90578-m. [DOI] [PubMed] [Google Scholar]
- Stewart P. L., Fuller S. D., Burnett R. M. Difference imaging of adenovirus: bridging the resolution gap between X-ray crystallography and electron microscopy. EMBO J. 1993 Jul;12(7):2589–2599. doi: 10.1002/j.1460-2075.1993.tb05919.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stouten P. F., Sander C., Ruigrok R. W., Cusack S. New triple-helical model for the shaft of the adenovirus fibre. J Mol Biol. 1992 Aug 20;226(4):1073–1084. doi: 10.1016/0022-2836(92)91053-r. [DOI] [PubMed] [Google Scholar]
- Valsesia-Wittmann S., Drynda A., Deléage G., Aumailley M., Heard J. M., Danos O., Verdier G., Cosset F. L. Modifications in the binding domain of avian retrovirus envelope protein to redirect the host range of retroviral vectors. J Virol. 1994 Jul;68(7):4609–4619. doi: 10.1128/jvi.68.7.4609-4619.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickham T. J., Mathias P., Cheresh D. A., Nemerow G. R. Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 1993 Apr 23;73(2):309–319. doi: 10.1016/0092-8674(93)90231-e. [DOI] [PubMed] [Google Scholar]