The sequence of adenovirus fiber: Similarities and differences between serotypes 2 and 5

doi:10.1016/0042-6822(87)90150-4

Virology

Volume 161, Issue 2, December 1987, Pages 549-554

https://doi.org/10.1016/0042-6822(87)90150-4 Get rights and content

Abstract

The amino acid sequence of the fiber from adenovirus type 5 has been deduced from the nucleotide sequence of the fiber gene. This sequence is compared with that of adenovirus type 2, a closely related serotype. We find 69% homology for the fiber protein whereas the known nonstructural proteins of these two serotypes have 99% sequence homology. A detailed sequence analysis was performed in the context of the model proposed by Green et al., [(1983), EMBO J., 8, 1357–1365] for the folding of the polypeptide chain of the adenovirus type 2 fiber. The N-terminal region, which in the virion is associated with the capsid, is identical for these two serotypes. In the shaft of the fiber the features, such as periodicity of the prolines and of the hydrophobic residues in the amino acid sequence, on which the model for the adenovirus type 2 is based are very well preserved in adenovirus type 5. On the other hand, there are large differences all along the sequence of the shaft of the fiber showing that there is a very limited homology between the amino acids in the two serotypes when they do not have a key role in establishing the structure. In the knob the homology between serotypes is 64%. These results are consistent with the differences between serotypes being confined to the exposed proteins.

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Cited by (61)

Efficient gene transfer into hematopoietic cells by a retargeting adenoviral vector system with a chimeric fiber of adenovirus serotype 5 and 11p
2006, Experimental Hematology
Citation Excerpt :
Finally, pFiber5/11p was digested with EcoRI, dephosphorized, and ligated with the preserved 23825 bp part of AdEasy-1 to generate pAdEasy-1/F11p. In the pAdEasy-1/F11p, the Ad5 sequence from nt 31169 to 32770 (Genbank accession no. AY339865), encoding 45-578 aa of Ad5 fiber, was replaced by the Ad11p sequence from nt 30940 to 31788 (Genbank accession no. AY598970), encoding 44-325 aa of Ad11p Fiber [2,20,21]. Codons of Ad5 fiber 579-581 aa in the polyadenylation signal were saved in pAdEasy-1/F11p, and they wouldn't be translated because the translation stop codon of Ad11p fiber was added just ahead of them.
Adenoviral vectors (Ad) were widely used in gene therapy and study of gene function, but the commonly used serotype 5 adenovirus-based vectors (Ad5) could poorly transduce hematopoietic cells because of low expression of viral receptors on these cells. To overcome this limitation, we developed a retargeting adenovector with a chimeric fiber of Ad5 and Ad11p (Ad5F11p) and evaluated its gene transfer ability in hematopoietic cells.
An Ad11p fiber pseudotyped Ad5 vector was generated by modifying the fiber gene of pAdEasy-1 backbone plasmid. Ad5F11p-GFP encoding enhanced green fluorescence protein (GFP) gene was transferred into human leukemic cell lines, primary leukemic cells, and CD34⁺ hematopoietic cells. The gene transduction efficiency was determined by fluorescence-activated cell sorting assay.
More than 90% of U937 or K562 cells could be infected by Ad5F11p-GFP at a moderate multiplicity of infection (MOI). Ad5F11p-GFP is also significantly more effective than control Ad5-GFP in infection of primary myeloid leukemic cells. At 200 MOI, GFP-positive percentages of Ad5F11p-GFP transduced myeloid leukemic cells range from 10.58% to 92.63% with a median of 28.65%. Ad5F11p-GFP could transduce about 50% human hematopoietic stem/progenitor (CD34⁺) cells, while Ad5-GFP could transduce <15% at 200 MOI. CD46 was reported to be the receptor of Ad11p. Our data suggest that CD46 participates in the process of Ad5F11p-GFP infection but is not the unique molecule determining its gene transfer efficiency of host cells.
We established a retargeting adenovector system, which could infect hematopoietic cells effectively and would benefit research work on Ad tropism.
Using GFP-ligand fusions to measure receptor-mediated endocytosis in living cells
2002, Vitamins and Hormones
Recombinant DNA technology has enabled the production of many types of chimeric proteins containing heterologous functional domains that have served a variety of useful capacities for cell biology research. Among proteins gaining wide use as a fusion partner is Aequorea victoria green fluorescent protein (GFP). GFP has been employed by numerous groups as a reporter gene for cell transfection and as an autofluorescent tag by recombinant fusion to foreign sequences. Here we describe the use of GFP as a tag for ligands, and provide examples of how purified recombinant GFP-ligand fusion proteins may be used to detect ligand-receptor interactions, including receptor-mediated endocytosis. Both its utility and limitations are discussed.
Mapping of linear epitopes on fibre knob of human adenovirus serotype 5
2001, Virus Research
Linear antigenic epitopes on the Ad5 fibre knob (FK5) were characterised with fibre- and virion-specific antisera, using 15-mer overlapping peptides covering the knob of the fibre. They were compared with epitopes on the Ad2 fibre knob (FK2) domain. The stronger reactive FK5 epitopes were represented by peptides P3 (amino acids (aa A419–L433)), P6 (aa S449–E463), P7 (aa I459–L473), P12 (aa G509–N523), P14 (aa P529–G543) and P16 (aa A549–Y563). P3 spans the B β-strand and the left portion of the C β-strand, P6 and P7 the D β-strand and the adjacent parts of the CD and DE loops, P12, P14 and P16 the G, H and I β strands and the adjacent parts of the loops, respectively. The stronger reactive epitopes on FK2 were located in P2 (aa P409–L423), P6 (aa T449–Q463), P8 (aa E469–G483), P13 (aa Q519–T533) and P16 (aa S549–K563). The positions of FK5 and FK2 derived peptides, representing epitopes, are either identical or overlapping or adjacent, as determined by amino acid sequence alignment. Antisera obtained against several longer peptides showed virus neutralising capacity, indicating neutralising epitopes in these peptides.
Specific subgroup B adenovirus diagnosis by PCR of the fibre gene
2000, Journal of Infection
Objective: A highly sensitive and specific PCR assay targeting regions of the fibre gene was developed for the identification of subgroup B adenovirus strains. This is critical, since these adenovirus strains are frequently associated with severe respiratory infections in infants and new-borns.
Methods: Clinical samples from nasopharyngeal aspirates were analysed by PCR using several sets of primers corresponding to sequences of the gene coding for the fibre protein.
Results: The assay allowed the detection and identification of all the genotypes of adenovirus subgroup B, based on the size of the amplified product when analysed on polyacrilamide gel electrophoresis. Specifically, one set of primers was able to amplify DNA of subgroup B but not subgroup C and E viruses.
Conclusion: The detection of adenovirus and the genotyping can be done on a routine basis by a PCR assay using the fibre gene as a target. The assay allows the identification of ADV subgroup B, including genotype 7h, which is the single most important viral pathogen associated with respiratory diseases in infants and young children in the southern part of South America.
Two closely related adenovirus genome types with kidney or respiratory tract tropism differ in their binding to epithelial cells of various origins
1998, Virology
The host–cell interactions of the genome types Ad11p and Ad11a of human adenovirus serotype 11, displaying kidney or respiratory tropism, were compared using FACS analysis. Kinetic experiments indicated that the virus binding started immediately and reached a plateau after 30 min. The binding of biotinylated virions to seven continuous cell lines: A549, A498, J82, HeLa, CHO, MDCK, and human diploid fibroblasts (HEDF), was quantitated by FACS analysis. The binding capacities of the two viruses to all human cell lines but A549 cells appeared to differ. Ad11p virions manifested high affinities, whereas Ad11a virions presented low affinities. Neither of the two viruses bound to CHO or MDCK cells. Reciprocal competition experiments showed that the Ad11a virions could be weakly blocked by the Ad11p virions, whereas the Ad11p virions could not be competed at all by the Ad11a virions. The binding of the Ad11p virions to cells could be blocked by the rfiber antiserum of Ad11p, but not by the corresponding antiserum against Ad11a or Ad35p. A comparison of the cytopathogenicity of the seven cell lines infected by Ad11p and Ad11a demonstrated that the efficiency of the initial event of an adenovirus infection directly affects the outcome of the viral infection. The Ad11a in the A498, J82, HeLa, or HEDF cells that presented lower affinity and receptor concentration showed 100 times less infectivity than that in A549 cells displaying high affinity and receptor concentration. These results indicate that the cell susceptibility to Ad11p and Ad11a infection strongly depends on both the number of fiber receptors on the host cells and the receptor affinity for ligands on the fiber knob. Our findings also suggest that the receptors for Ad11p and Ad11a on the surface of different cell types may be different or on different sites.
Inhibition of cell adhesion to the virus by synthetic peptides of fiber knob of human adenovirus serotypes 2 and 3 and virus neutralisation by anti-peptide antibodies
1996, Virus Research
The fiber knob of adenovirus (Ad) causes the first step in the interaction of adenovirus with cell membrane receptors. To obtain information on the receptor binding site(s) several synthetic peptides derived from Ad2 and Ad3 fiber head sequences and their antisera were tested for interference with virus attachment to HeLa and FL cells and cell adhesion to viruses. The anti-peptide sera were also evaluated in ELISA and virus neutralisation test. Ad2 (of subgroup C) and Ad3 (of subgroup B) attachment was not significantly inhibited by peptides corresponding to the amino acid residues 535–554, 555–573, 562–582 of Ad2 fiber or 210–225, 267–283, 291–306 and 300–319 of Ad3 fiber. However, microplate pre-adsorbed Ad3 fiber residues 210–225 and 267–283 could bind FL and HeLa cells, and 1 mg/ml of Ad3 fiber residues 267–283 inhibited the cell adhesion to Ad3 virus to approximately 90%. This peptide may participate in the receptor binding site of Ad3 fiber. ELISA reactive anti-peptide antibodies against the homologous peptide and virus did not significantly reduce the cell adhesion to the immobilised virus or the virus attachment to cells, but in the neutralisation assay antibodies raised to Ad2 fiber residues 555–573 and 562–582 and Ad3 fiber residues 210–225 caused neutralisation of the homologous virus at serum dilutions of 1:500 and 1:32, respectively. The corresponding peptides and one further peptide of Ad2 fiber and two of Ad3 fiber seem to contain neutralisation epitopes.

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Virology

Abstract

References (23)

The molecular basis of antigenic variation in influenza virus

Adv. Virus Res.

DNA sequence of the early E3 transcription unit of adenovirus 5

Virology

Crystal packing and stoichiometry of the fibre protein of adenovirus type 2

J. Mol. Biol.

Restriction maps of human adenovirus types 2, 5 and 3 for Bcll, Clal, Pvul and Sphl endonucleases

Gene

Nucleotide sequence from the adenovirus-2 genome

J. Biol. Chem.

Thirty-one human adenovirus serotypes (Adl–Ad31) form five groups (A–E) based upon DNA genome homologies

Virology

Adenovirus hexon. Sequence comparison of subgroup C serotypes 2 and 5

J. Biol. Chem.

Restriction maps of human adenovirus types 2, 5 and 3 for BstEll, Mlul, Ndel, Nrul and Sfil endonucleases

Gene

Complete nucleotide sequences of all three poliovirus serotypes genomes

J. Mol. Biol.

Rapid,and easy sequencing of large linear doubled-stranded DNA and supercoiled plasmid DNA

Gene Anal. Tech.

Virology

Cited by (61)

Efficient gene transfer into hematopoietic cells by a retargeting adenoviral vector system with a chimeric fiber of adenovirus serotype 5 and 11p

Using GFP-ligand fusions to measure receptor-mediated endocytosis in living cells

Mapping of linear epitopes on fibre knob of human adenovirus serotype 5

Specific subgroup B adenovirus diagnosis by PCR of the fibre gene

Two closely related adenovirus genome types with kidney or respiratory tract tropism differ in their binding to epithelial cells of various origins

Inhibition of cell adhesion to the virus by synthetic peptides of fiber knob of human adenovirus serotypes 2 and 3 and virus neutralisation by anti-peptide antibodies