Adenovirus type 12-induced rat tumor cells of neuroepithelial origin: persistence and expression of the viral genome

Ibelgaufts, Horst; Doerfler, Walter; Scheidtmann, Karl Heinz; Wechsler, W.

doi:10.1128/jvi.33.1.423-437.1980

Cited by 39 publications

(9 citation statements)

References 42 publications

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“…In some of the Adl2transformed hamster lines, only early genes are transcribed into mRNA (22). Early and late genes are expressed in Adl2-induced rat brain tumor cell lines (19). However, expression of late genes in adenovirus-transformed cells appears to be the exception rather than the rule.…”

Section: Giessen2 Germanymentioning

confidence: 99%

“…There is now very good evidence for the integrated state of adenovirus DNA in a number of different systems (for review, see reference 6). In most adenovirus type 12 (Adl2)-transformed cell lines or Adl2-induced tumor cell lines, the entire viral genome persists in multiple copies (8, 10, 14, 20) in a very stably integrated form (8,15,16,19,33). However, in many of the Ad2and Ad5-transformed rat cell lines investigated, only fractions of the viral genome are present (26) in an integrated form (12).…”

Section: Giessen2 Germanymentioning

confidence: 99%

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Integration Sites of Adenovirus Type 12 DNA in Transformed Hamster Cells and Hamster Tumor Cells

Stabel

Doerfler

Friis

1980

J Virol

114

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Preparation ofthe E. coli plasmid pBR322 (30). The plasmid pBR322 was prepared as described earlier (8). Extraction and purification of cellular DNA. The sodium dodecyl sulfate (SDS)-pronase B-phenol procedure was outlined in detail elsewhere (33). The DNA was also treated with RNase and proteinase K and was purified by equilibrium centrifugation in CsCl density gradients (33). Analysis of cellular DNA by the Southern blotting technique (29). The total intracellular DNA from transformed cells was cleaved with various re

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Section: Giessen2 Germanymentioning

confidence: 99%

Section: Giessen2 Germanymentioning

confidence: 99%

Integration Sites of Adenovirus Type 12 DNA in Transformed Hamster Cells and Hamster Tumor Cells

Stabel

Doerfler

Friis

1980

J Virol

114

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“…DISCUSSION In each of the cell lines or tumor lines investigated, a characteristic set of Adl2-specific RNA molecules is synthesized. These RNA molecules are derived mainly from the early segments of the Ad12 genome; in some cell lines (CLAC3 and RBT12/3), cytoplasmic RNA is also derived from late segments (13). The data compiled in Table 1 summarize all of the results obtained and list the size classes of Adl2-specific RNAs identified in the cytoplasmic RNA of each cell line.…”

Section: Survey Of Cell Lines Investigated Formentioning

confidence: 99%

“…Cell lines HA12/7 (32) and A2497-3 (A. M. Lewis, Jr., personal communication) have been derived from primary hamster cells by transformation with Adl2 in culture. Lines CLAC3 (22) and RBT12/3 (13) have been established from an Adl2-induced subcutaneous hamster tumor and an Adl2-induced rat brain tumor, respectively. The hamster cell line B3 was used as a control in all experiments.…”

Section: Survey Of Cell Lines Investigated Formentioning

confidence: 99%

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Expression of Viral DNA in Adenovirus Type 12-Transformed Cells, in Tumor Cells, and in Revertants

Schirm

Doerfler

1981

J Virol

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The expression as cytoplasmic RNA of integrated human adenovirus type 12 (Ad12) DNA in transformed and tumor cell lines and in revertants was investigated. The transformed and tumor cells contained multiple copies of the viral genome, 3 to 22 copies per cell in different cell lines. The integrated Ad12 DNA molecules persisted intact or nearly intact and in most cases colinear with the virion DNA. In the revertant cell lines, which were derived from cell line T637 (22 copies of Ad12 DNA per cell), all of the Ad12 DNA molecules were lost (line F10) or only one copy and a fraction of a second copy persisted (line TR12). The size classes and map locations of Ad12-specific cytoplasmic RNAs in three Ad12-transformed hamster cell lines (T637, HA12/7, and A2497-3), in two revertant lines (F10 and TR12), in one Ad12-induced hamster (CLAC3), and in one rat brain tumor line (RBT12/3) were determined. Cytoplasmic RNA from uninfected B3 hamster cells and from human KB cells productively infected with Ad12 served as controls. In the latter control experiments, the RNA was isolated early or late postinfection. With respect to the amounts of Ad12-specific RNAs detected in cytoplasmic RNA from various Ad12-transformed or Ad12-induced tumor cell lines, we could not establish any correlations to the number of Ad12 genome copies integrated into the cellular DNAs. Thus, the expression of the integrated viral genomes in these lines was regulated by mechanisms more complicated than simple gene dosage effects. Using cloned fragments of Ad12 DNA as hybridization probes, we analyzed the cytoplasmic RNAs from the cell lines mentioned by electrophoresis on agarose gels, blotting, and DNA-RNA hybridization. For each transformed and tumor cell line, except for the revertants, several size classes of Ad12-specific cytoplasmic RNA were detected for the early E1, E2, and E4 regions of Ad12 DNA. Some of these size classes were similar but not identical to those observed in cytoplasmic RNA isolated early from human KB cells productively infected with Ad12. Only cell lines A2497-3, T637, and RBT12/3 contained several size classes of cytoplasmic RNA homologous to the E3 region of Ad12 DNA. Weak homologies to the E1 region of Ad12 DNA were also detected in the revertant lines F10 and TR12. Late regions of Ad12 DNA were expressed as cytoplasmic RNA in cell lines CLAC3 and RBT12/3. Weak homologies were detected between certain segments of the Ad12 genome (the Eco RI-B, -C, and -D fragments) and the cytoplasmic RNA from uninfected hamster cells. These homologies had no apparent counterpart at the level of DNA, perhaps because these homologies could be detected only due to an overrepresentation of RNA sequences. In preliminary experiments, we failed to detect the expression as cytoplasmic RNA of the so-called virus-associated RNA in transformed and tumor cell lines. Virus-associated RNA represents a population of low-molecular-weight RNAs that map at around 30 fractional length units on the viral genome.

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Covalently closed circles of human adenovirus DNA are infectious.

Fl¹

1984

The EMBO Journal

100

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Replication of the linear adenovirus DNA molecule is thought to result from semiconservative synthesis off linear templates, starting from origins at either end of the genome. Recently, however, it has been shown that in cells infected with adenovirus type 5 (Ad5) a significant fraction of the ends of viral DNA molecules become joined head‐to‐tail due at least in part to the formation of covalently closed circles. Circular DNA is not present in virions but joining of the ends of viral DNA is detectable shortly after infection, well before the onset of viral DNA replication. To learn more about the structure and possible function of these circular forms of viral DNA, I have cloned Ad5 circles as plasmids replicating in Escherichia coli. Two plasmids have been analyzed in detail and shown to generate infectious virus with an efficiency comparable with that of virion DNA following transfection into human cells. These results suggest that circles are not totally inert or functionless but that, once formed, they are capable of re‐entering the pool of replicating molecules to generate linear progeny.

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Adenovirus type 12-induced rat tumor cells of neuroepithelial origin: persistence and expression of the viral genome

Cited by 39 publications

References 42 publications

Integration Sites of Adenovirus Type 12 DNA in Transformed Hamster Cells and Hamster Tumor Cells

Integration Sites of Adenovirus Type 12 DNA in Transformed Hamster Cells and Hamster Tumor Cells

Expression of Viral DNA in Adenovirus Type 12-Transformed Cells, in Tumor Cells, and in Revertants

Covalently closed circles of human adenovirus DNA are infectious.

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