The ultrastructure and nature of adenovirus type 2-induced paracrystalline formations

Henry, Claude J.; Slifkin, Malcolm; Merkow, Leonard P.; Pardo, Matias

doi:10.1016/0042-6822(71)90167-x

Cited by 14 publications

(6 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In line with this hypothesis, previous investigators provided structural description of the crystals similar to that presented in this study, and morever, by using temperature-sensitive ( ts ) mutants, suggested that the crystals were derived from fibers, with a proviso that penton base might also be involved [28]. However, other studies suggested that the crystals were formed of basic core proteins [23], [24], [26]. More recent immuno-EM analyses revealed that the intranuclear crystals reacted with anti-hexon, anti-penton base and anti-fiber knob antibodies [43], as well as with protein kinase CK2 and dsRNA-activated protein kinase PKR antibodies, leading to the assumption that multiple viral and cellular proteins were components of the crystals [44].…”

Section: Discussionsupporting

confidence: 82%

Protein Crystals in Adenovirus Type 5-Infected Cells: Requirements for Intranuclear Crystallogenesis, Structural and Functional Analysis

et al. 2008

View full text Add to dashboard Cite

Intranuclear crystalline inclusions have been observed in the nucleus of epithelial cells infected with Adenovirus serotype 5 (Ad5) at late steps of the virus life cycle. Using immuno-electron microscopy and confocal microscopy of cells infected with various Ad5 recombinants modified in their penton base or fiber domains, we found that these inclusions represented crystals of penton capsomers, the heteromeric capsid protein formed of penton base and fiber subunits. The occurrence of protein crystals within the nucleus of infected cells required the integrity of the fiber knob and part of the shaft domain. In the knob domain, the region overlapping residues 489–492 in the FG loop was found to be essential for crystal formation. In the shaft, a large deletion of repeats 4 to 16 had no detrimental effect on crystal inclusions, whereas deletion of repeats 8 to 21 abolished crystal formation without altering the level of fiber protein expression. This suggested a crucial role of the five penultimate repeats in the crystallisation process. Chimeric pentons made of Ad5 penton base and fiber domains from different serotypes were analyzed with respect to crystal formation. No crystal was found when fiber consisted of shaft (S) from Ad5 and knob (K) from Ad3 (heterotypic S5-K3 fiber), but occurred with homotypic S3K3 fiber. However, less regular crystals were observed with homotypic S35-K35 fiber. TB5, a monoclonal antibody directed against the Ad5 fiber knob was found by immunofluorescence microscopy to react with high efficiency with the intranuclear protein crystals in situ. Data obtained with Ad fiber mutants indicated that the absence of crystalline inclusions correlated with a lower infectivity and/or lower yields of virus progeny, suggesting that the protein crystals might be involved in virion assembly. Thus, we propose that TB5 staining of Ad-infected 293 cells can be used as a prognostic assay for the viability and productivity of fiber-modified Ad5 vectors.

show abstract

Section: Discussionsupporting

confidence: 82%

Protein Crystals in Adenovirus Type 5-Infected Cells: Requirements for Intranuclear Crystallogenesis, Structural and Functional Analysis

et al. 2008

View full text Add to dashboard Cite

show abstract

“…It can be predicted that the study of adenovirus will also benefit from the latest advances in the EM field. Crystalline inclusions have been reported in cell nuclei at late stages of infection (BOULANGER et al 1970;HENRY et al 1971). 8) Finally, it is also expected that the adenovirus particle will eventually yield to analysis by X-ray crystallography.…”

Section: Discussionmentioning

confidence: 99%

Structural Studies on Adenoviruses

Martı́n

Burnett

2003

Current Topics in Microbiology and Immunology

View full text Add to dashboard Cite

“…Positive fluorescence of paracrystals with phenanthrenequinone, a compound that forms a fluorescent condensate with arginine, suggests that they are composed of basic proteins, a characteristic common to core proteins. Originally, by ultrastructure observations, the Ad2-induced paracrystals (14) were thought to be composed of hexon, penton, and fiber antigens arranged in a small hexagonal structure as described previously by Boulanger et al (5). However, in this study, hexon antiserum prepared against AdS failed to stain Ad2-induced paracrystals.…”

Section: Discussionmentioning

confidence: 46%

Paracrystal Formation in Cell Cultures Infected with Adenovirus Type 2

Henry

Atchison

1971

J Virol

Self Cite

View full text Add to dashboard Cite

Large rod-shaped structures corresponding to paracrystals were seen in the nucleus, cytoplasm, or both of adenovirus type 2 (Ad2)-infected cells by immunofluorescence staining with antibody prepared against purified Ad2. In exception to this, Ad2-induced crystals did not stain with either hexon or fiber antibody. The crystalline structures were first observed in Ad2-infected Vero cells at 28 hr with a maximum number at 70 hr postinoculation. The kinetics of paracrystalline formation closely paralleled the experimental synthesis of infectious progeny virus. Acridine-orange staining revealed the lack of nucleic acids associated with the crystal. Also, the paracrystals stained intensely with phenanthrenequinone, suggesting that they are composed of basic proteins. Interferon induced by Newcastle disease virus from African green monkey kidney cell cultures was used to pretreat Vero cells prior to Ad2 infection. This resulted in inhibiting the formation of viral-induced paracrystals in 97% of the cells and reduced virus yields by 95%. The African green monkey kidney cell culture interferon did not reduce Ad2 yields in HeLa cell cultures or display any virus inhibitory activity in rabbit kidney cell cultures. Staining procedures, fluorescent-antibody tests with whole virus, hexon or fiber antibody, and interferon studies suggested that the paracrystals were viral-directed and composed of basic proteins (possibly core proteins).

show abstract

The ultrastructure and nature of adenovirus type 2-induced paracrystalline formations

Cited by 14 publications

References 12 publications

Protein Crystals in Adenovirus Type 5-Infected Cells: Requirements for Intranuclear Crystallogenesis, Structural and Functional Analysis

Protein Crystals in Adenovirus Type 5-Infected Cells: Requirements for Intranuclear Crystallogenesis, Structural and Functional Analysis

Structural Studies on Adenoviruses

Paracrystal Formation in Cell Cultures Infected with Adenovirus Type 2

Contact Info

Product

Resources

About