Localization of adenovirus DNA replication in KB cells

Vlak, J.M.; Rozijn, Th.H.; Spies, F.

doi:10.1016/0042-6822(75)90058-6

Cited by 18 publications

(11 citation statements)

References 34 publications

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“…The IF structures appeared randomly throughout the nucleoplasm, with no apparent association with the nuclear membrane. If DBP does function in viral DNA replication, this result is in accord with the reports that group C adenovirus DNA replication occurs throughout the nucleus and not in association with the nuclear membrane (36,41).…”

Section: Resultssupporting

confidence: 89%

Immunofluorescence Study of the Adenovirus Type 2 Single-Stranded DNA Binding Protein in Infected and Transformed Cells

Sugawara

Gilead

Wold

et al. 1977

J Virol

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High-titer monospecific antiserum against highly purified adenovirus 2 (Ad2) single-stranded DNA binding protein (DBP) was used to study, by indirect immunofluorescence (IF), the synthesis of DBP in Ad2-infected human cells and adenovirus-transformed rat, hamster, and human cell lines. In infected cells the synthesis of DBP was first detected in the cytoplasm at 2 to 4 h postinfection and reached a maximum intensity at 6 h postinfection. At this time DBP began to accumulate in the nucleus, where it reached maximum intensity at about 14 h postinfection. The cytoplasmic IF was diffuse, whereas nuclear IF appeared as dots that coalesced into large globules as infection progressed. In cells treated with 1-,3-D-arabinofuranosylcytosine to inhibit viral DNA synthesis, strong nuclear IF was observed in the form of dots, but the large fluorescent globules were not observed. The Ad2 (oncogenic group C) anti-DBP serum reacted very strongly by IF with Ad5 (group C)-infected, to a lesser extent with Ad7 and Adli (group B)-infected, and weakly with Ad12 and Adi8 (group A)-infected KB cells (treated with 1-,8-D-arabinofuranosylcytosine). These results may indicate that Ad2 DBP is closely related immunologically to DBPs induced early after infection by adenovirus serotypes in oncogenic group C, moderately related to DBPs of serotypes in oncogenic group B, and perhaps distantly related to DBPs of serotypes in oncogenic group A. The following adenovirus-transformed cell lines were examined for DBP synthesis by IF with the Ad2 anti-DBP serum: six rat cell lines (T2C4, F17, 8662, 8638, 8617, and F161) transformed by Ad2 virus, three hamster cell lines transformed by Ad2 virus (Ad2HT1) and Ad2-simian virus 40 hybrid virus (ND1HK1 and ND4HK4), and one rat (5RK) and one human (293-31) cell line transformed by transfection with Ad5 DNA. T2C4 and 8662 appeared weakly positive, whereas Ad2HT1 and ND4HK1 were strongly positive. The other transformed cell lines did not produce DBP detectable by IF. Thus, some but not all transformed cell lines produce DBP, which indicates that DBP is not required for maintenance of cell transformation and that transformed cells can express "nontransforming" viral genes as protein.Human adenoviruses are oncogenic DNA viruses that mature in the nucleus of permissive human cells and transform semipermissive or nonpermissive cell types with low frequency (reviewed in reference 31; W. S. M. Wold, M. Green, and W. Buttner, in D. P. Nayak [ed.], Molecular Biology of Animal Viruses, in press). There are at least 31 human adenovirus serotypes; some serotypes fall into three welldefined oncogenic groups: group A (adenovirus 12

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Section: Resultssupporting

confidence: 89%

Immunofluorescence Study of the Adenovirus Type 2 Single-Stranded DNA Binding Protein in Infected and Transformed Cells

Sugawara

Gilead

Wold

et al. 1977

J Virol

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“…As shown in Figure IA the distribution of Ad2 DNA within a HeLa cell nucleus is in multiple discrete foci or 'replication factories' which are not nucleoli (9)(10)(11)15,16). Figure 1A is a computer reconstructed composite of all optical sections and represents the total Ad2 DNA in a single HeLa cell nucleus.…”

Section: Intranuclear Localization Of Ad2 Dna In Hela Cellsmentioning

confidence: 99%

“…Our data on 3-D localization of Ad2 replication factories are consistent with both previous EM and M banding studies for intranuclear compartmentalization of adenovirus DNA replication and suggest a way that the apparently contradictory data can be reconciled. The EM data indicated that tritiated adenovirus DNA could be found in sections throughout the nucleus (9)(10)(11). We have seen that Ad2 replication factories were associated with the nuclear envelope, but often they were large and extended into the nuclear interior.…”

Section: Intranuclear Localization Of Ad2 Dna In Hela Cellsmentioning

confidence: 99%

“…Studies done during the 1970s on the intranuclear localization of adenovirus DNA gave ambiguous results. Studies using electron microscopy (EM) indicated that adenovirus DNA is replicated within the nucleus in multiple foci termed 'replication factories' and that tritiated adenoviral DNAs appear to be found throughout the entire nucleus with no preferential association with the nuclear envelope (9)(10)(11). However, biochemical studies using the 'M banding' technique indicated that adenovirus DNA replication is compartmentalized along the nuclear membrane (10)(11)(12).…”

Section: Introductionmentioning

confidence: 99%

“…These interactions are most likely mediated by the k./ 1990 Oxford University Press 513 terminal proteins covalently attached to the 5' ends of the linear viral DNAs (14). Both Ad2 and MVM DNA are compartmentalized spatially in that they are found in foci within infected cell nuclei; MVM DNA is localized in nucleoli of infected cells (15) while Ad2 is localized in multiple non-nucleolar 'replication factories' (9)(10)(11)15,16). The Ad2 replication factories contain several nucleolar antigens which are recompartmentalized during Ad2 infection, and when MVM is coinfected with Ad2 in HeLa cells its DNA and terminal protein are also recompartmentalized from nucleoli to the replication factories (15).…”

Section: Introductionmentioning

confidence: 99%

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Adenovirus and minute virus of mice DNAs are localized at the nuclear periphery

1990

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The localization of adenovirus 2 (Ad2) and Minute Virus of Mice (MVM) DNAs was studied in situ in infected HeLa cell nuclei using fluorescent DNA probes and confocal microscopy. Ad2 DNA was found in multiple foci which were localized along the periphery of the infected cell nuclei. MVM DNA was found in HeLa cell nucleoli which are associated with the nuclear envelope, and when co-infected with Ad2 MVM DNA was compartmentalized to multiple foci which again were localized at the nuclear periphery. The data are discussed in terms of a model for the role of intranuclear compartmentalization in eukaryotic DNA structure and function.

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Recent Trends in Electron Microscope Autoradiography

Maraldi

1982

Cell Growth

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Localization of adenovirus DNA replication in KB cells

Cited by 18 publications

References 34 publications

Immunofluorescence Study of the Adenovirus Type 2 Single-Stranded DNA Binding Protein in Infected and Transformed Cells

Immunofluorescence Study of the Adenovirus Type 2 Single-Stranded DNA Binding Protein in Infected and Transformed Cells

Adenovirus and minute virus of mice DNAs are localized at the nuclear periphery

Recent Trends in Electron Microscope Autoradiography

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