Defect of adenovirus type 12 replication in hamster cells: absence of transcription of viral virus-associated and L1 RNAs

Jüttermann, Ruth; Weyer, Ulrike; Doerfler, Walter

doi:10.1128/jvi.63.8.3535-3540.1989

Cited by 12 publications

(3 citation statements)

References 30 publications

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“…Among rodents, hamsters, and cotton rats have been proposed as suitable models for respiratory infection by human adenovirus (22,30,46). Hamster cells are permissive for Ad2 and Ad5 replication (35) but are significantly less permissive than human cells, while mouse cells are semipermissive (7,51). In this study, we found that porcine cell lines support Ad5 replication, showing virus kinetics and virus titers similar to those obtained for human epithelial cells.…”

Section: Discussionsupporting

confidence: 66%

Tropism of human adenovirus type 5-based vectors in swine and their ability to protect against transmissible gastroenteritis coronavirus

Torres

Alonso

Ortega

et al. 1996

J Virol

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The infection of epithelia] swine testicle and intestinal porcine epithelial (IPEC-1) cell lines by adenovirus type 5 (Ad5) has been studied in vitro by using an Ad5-luciferase recombinant containing the firefly luciferase gene as a reporter. Porcine cell lines supported Ad5 replication, showing virus titers, kinetics of virus production, and luciferase expression levels similar to those obtained in human 293 cells, which constitutively express the 5'-end 11% of the Ad5 genome. The tropism of Ad5-based vectors in swine and its ability to induce an efficient immune response against heterologous antigens expressed by foreign genes inserted in these vectors has been determined. Ad5 vectors replicate and express heterologous antigens in porcine lungs and mediastinal and mesenteric lymph nodes. Significant levels of heterologous antigen expression were also demonstrated in the small intestine (jejunum and ileum), but Ad5 replication in this organ was very poor, suggesting that Ad vectors undergo an abortive replication in the porcine small intestine. The tissues infected by Ad5 were dependent on the inoculation route. The oronasal route appeared to be best for inoculation of bronchus-associated lymphoid tissue infection, while the intraperitoneal route was best for gut-associated lymphoid tissue infection. Epithelial cells of bronchioles, macrophages, type II pneumocytes, and follicular dendritic cells were identified as targets for Ad5, while epithelial cells of the intestine were not infected by Ad5. Viruses with a deletion from 79.5 to 84.8 map units in the E3 region, with or without heterologous inserted genes, replicated to lower levels in porcine tissues than did wild-type Ad5. It was also shown that an Ad5 recombinant expressing the four antigenic sites (A, B, C, and D) of transmissible gastroenteritis coronavirus (TGEV) spike protein induced in swine immune responses which neutralized TGEV infectivity. In addition, porcine serum from Ad-TGEV-immune animals provide passive protection when mixed with fully virulent TGEV and orally administered to highly susceptible newborn piglets. These results taken together indicate that swine may be a good animal model for human Ad5 lung infection to aid in the evaluation of candidate adenovirus vaccines and that Ad5 may be suitable as a recombinant viral vaccine or for other applications in swine.

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Section: Discussionsupporting

confidence: 66%

Tropism of human adenovirus type 5-based vectors in swine and their ability to protect against transmissible gastroenteritis coronavirus

Torres

Alonso

Ortega

et al. 1996

J Virol

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“…It seemed likely that there were several defects in the Adl2 genome to account for the abortive cycle of Adl2 in hamster cells. In previously published analyses, these defects had been localized to the E1B region (Klimkait and Doerfler, 1987), to the VA RNA region (Juttermann et al, 1989), and to the MLP of Adl2 DNA (Weyer and Doerfler, 1985). It was conceivable that these functions were somehow interdependent since all were subject to ElA regulation.…”

Section: Resultsmentioning

confidence: 99%

A mitigator sequence in the downstream region of the major late promoter of adenovirus type 12 DNA.

Zock

Doerfler

1990

The EMBO Journal

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Human adenovirus type 12 (Ad12) replicates in permissive human host cells, but undergoes an abortive infection cycle in non‐permissive hamster cells. Ad12 DNA cannot replicate and late viral genes are not expressed in hamster cells, whereas most of the early viral mRNAs are synthesized. We have shown previously that the major late promoter of Ad12 DNA (Ad12 MLP; nucleotides ‐228 to +435 relative to nucleotide +1 as the site of transcriptional initiation) does not function in uninfected or in Ad12‐infected hamster BHK21 cells. The transcriptional defect of Ad12 DNA in hamster cells has thus been, at least partly, localized to the viral MLP. As expected, this construct is active in permissive human cells. Here, we show that the sequence between nucleotides +249 and +435 in the Ad12 MLP is in some way responsible for the late transcriptional block of this promoter in hamster cells. An Ad12 MLP‐‐CAT construct comprising nucleotides ‐228 to +248 shows striking activity in hamster cells, and its activity is very markedly enhanced in Ad2‐ or Ad12‐infected hamster or human cells compared with the nucleotide ‐228 to +435 construct. By using exonuclease Bal31, a series of Ad12 MLP‐‐CAT gene assemblies were constructed which carry deletions of increasing lengths in the downstream part of the Ad12 MLP. Activity measurements of these constructs in BHK21 and in HeLa cells have located the presumptive mitigator element to the Ad12 sequence between nucleotides +320 and +352 of the MLP. It is also demonstrated that in the nucleotide ‐228 to +248 MLP construct, transcription is initiated at the authentic Ad12 MLP cap site after the transfection of both hamster and human cells. The localization of this cap site in the nucleotide sequence of the Ad12 MLP indicates the similarity to the comparable start site in the MLP of Ad2 DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…The levels of Adl2 ElA and E1B gene products in BHK21 cell extracts have been reported to be about 10% of those in human KB cell extracts (33). Late Adl2 genes or the virus-associated RNA region encoding virus-associated RNA are not transcribed in hamster cells (23). The block of Adl2 DNA replication and late-gene transcription in hamster * Corresponding author.…”

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confidence: 99%

A unique mitigator sequence determines the species specificity of the major late promoter in adenovirus type 12 DNA

Zock

Iselt

Doerfler

1993

J Virol

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Human adenovirus type 12 (Adl2) cannot replicate in hamster cells, whereas human cells are permissive for Adl2. Adl2 DNA replication and late-gene and virus-associated RNA expression are blocked in hamster cells. Early Adl2 genes are transcribed, and the viral DNA can be integrated into the host genome. Adl2 DNA replication and late-gene transcription can be complemented in hamster cells by El functions of Ad2 or Ad5, for which hamster cells are fully permissive (for a review, see W. Doerfler, Adv. Virus Res. 39:89-128, 1991). We have previously demonstrated that a 33-nucleotide mitigator sequence, which is located in the downstream region of the major late promoter (MLP) of Adl2 DNA, is responsible for the inactivity of the Adl2 MLP in hamster cells (C. Zock and W. Doerfler, EMBO J. 9:1615-1623, 1990). A similar negative regulator has not been found in the MLP of Ad2 DNA. We have now studied the mechanism of action of this mitigator element. The results of nuclear run-on experiments document the absence of MLP transcripts in the nuclei of Ad12-infected BHK21 hamster cells. Surprisingly, the mitigator element cannot elicit its function in in vitro transcription experiments with nuclear extracts from both hamster BHK21 and human HeLa cells. Intact nuclear topology and/or tightly bound nuclear elements that cannot be eluted in nuclear extracts are somehow required for recognition of the Adl2 mitigator. Electrophoretic mobility shift assays have not revealed significant differences in the binding of proteins from human HeLa or hamster BHK21 cells to the mitigator sequence in the MLP of Adl2 DNA or to the corresponding sequence in Ad2 DNA. We have converted the sequence of the mitigator in the MLP of Adl2 DNA to the equivalent sequence in the MLP of Ad2 DNA by site-directed mutagenesis. This construct was not active in hamster cells. When the Adl2 mitigator, on the other hand, was inserted into the Ad2 MLP, the latter's function in hamster cells was not compromised. Deletions in the 5' upstream region of the Adl2 MLP have provided evidence for the existence of additional sequences that codetermine the deficiency of the Adl2 MLP in hamster cells. The amphifunctional YY1 protein

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Defect of adenovirus type 12 replication in hamster cells: absence of transcription of viral virus-associated and L1 RNAs

Cited by 12 publications

References 30 publications

Tropism of human adenovirus type 5-based vectors in swine and their ability to protect against transmissible gastroenteritis coronavirus

Tropism of human adenovirus type 5-based vectors in swine and their ability to protect against transmissible gastroenteritis coronavirus

A mitigator sequence in the downstream region of the major late promoter of adenovirus type 12 DNA.

A unique mitigator sequence determines the species specificity of the major late promoter in adenovirus type 12 DNA

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