Adenovirus Genetics

Williams, Jim

doi:10.1007/978-1-4613-2293-1_8

Cited by 18 publications

(4 citation statements)

References 262 publications

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“…(A) Result of screening several clones (derived from experiment 2 of Table 1 E1B, hr6, was unable to rescue AAV DNA replication in latently infected Detroit 6 cells (40), the ability of this mutant to rescue AAV DNA replication in the latently infected KB cells was tested. hr6-infected cells do not synthesize the E1B 55,000-molecular-weight protein (55K protein), although they do express the Ad5 E1B 21K protein (22,56). hr6 was able, as expected, to help AAV DNA replication in KB and 293 cells coinfected with hr6 and AAV ( Table 2).…”

Section: Resultsmentioning

confidence: 61%

“…In contrast to the previous report, however, hr6 did unquestionably rescue AAV DNA replication in all the latently infected KB cell clones, although it was less efficient than were the other helper viruses tested. Two other DNA-positive adenovirus mutants in the adenovirus E1B region, hrl3 (26,56) and Ad5sub316 (30), also rescued AAV DNA replication in the latently infected KB cells. A representative autoradiogram is shown in Fig.…”

Section: Resultsmentioning

confidence: 94%

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Latent infection of KB cells with adeno-associated virus type 2

Laughlin¹,

Cardellichio²,

Coon³

1986

J Virol

102

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Adeno-associated virus (AAV) is a prevalent human virus whose replication requires factors provided by a coinfecting helper virus. AAV can establish latent infections in vitro by integration of the AAV genome into cellular DNA. To study the process of integration as well as the rescue of AAV replication in latently infected cells after superinfection with a helper virus, we established a panel of independently derived latently infected cell clones. KB cells were infected with a high multiplicity of AAV in the absence of helper virus, cloned, and passaged to dilute out input AAV genomes. AAV DNA replication and protein synthesis were rescued from more than 10% of the KB cell clones after superinfection with adenovirus type 5 (Ad5) or herpes simplex virus types 1 or 2. In the absence of helper virus, there was no detectable expression of AAV-specific RNA or proteins in the latently infected cell clones. AdS superinfection also resulted in the production of infectious AAV in most cases. All mutant adenoviruses tested that were able to help AAV DNA replication in a coinfection were also able to rescue AAV from the latently infected cells, although one mutant, Ad5hr6, was less efficient at AAV rescue. Analysis of high-molecular-weight cellular DNA indicated that AAV sequences were integrated into the cell genome. The restriction enzyme digestion patterns of the cellular DNA were consistent with colinear integration of the AAV genome, with the viral termini present at the cell-virus junction. In addition, many of the cell lines appeared to contain head-to-tail concatemers of the AAV genome. The understanding of the integration of AAV DNA is increasingly important since AAV-based vectors have many advantages for gene transduction in vitro and in vivo.

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Section: Resultsmentioning

confidence: 61%

Section: Resultsmentioning

confidence: 94%

Latent infection of KB cells with adeno-associated virus type 2

Laughlin¹,

Cardellichio²,

Coon³

1986

J Virol

102

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“…How these modes of positive and negative regulation by ElA may be involved in ElA-dependent viral transformation and tumorigenesis (reviewed in ref. 8) is unknown.…”

mentioning

confidence: 99%

The 289-amino acid E1A protein of adenovirus binds zinc in a region that is important for trans-activation.

Culp¹,

Webster²,

Friedman³

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

102

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The ElA gene of adenovirus type 5 encodes two major proteins of 289 and 243 amino acid residues, which are identical except that the larger protein has an internal stretch of 46 amino acids required for efficient trans-activation of early viral promoters. This domain contains a consensus zinc finger motif (Cys-Xaa2-Cys-Xaal3-Cys-Xaa2-Cys) in which the cysteine residues serve as postulated ligands. Atomic absorption spectrophotometry applied to bacterially expressed ElA proteins revealed that the 289-amino acid protein binds one zinc ion, whereas the 243-amino acid protein binds no zinc. Replacing individual cysteine residues of the finger with other amino acids destroyed the trans-activating ability of the 289-amino acid protein, even when structurally or functionally conserved amino acids were substituted. These results strongly suggest that the zinc finger of the 46-amino acid domain is intimately linked to the ability of the large EMA protein to stimulate transcription of ElA-inducible promoters. Furthermore, zinc binding to one of the mutant finger proteins suggests either that only a precise finger structure formed by the tetrahedral coordination of zinc to the four consensus ligands is required for trans-activation or, possibly, that one of several neighboring histidine residues in various combinations with three of the consensus cysteine residues normally coordinates zinc. How the zinc finger in ElA might interact with DNA or protein to bring about trans-activation is discussed.The ElA gene ofadenovirus functions normally by transcriptionally activating (trans-activating) other early viral promoters, which leads to productive infection (1, 2). ElA gene products can also trans-activate the promoters of some cellular genes-e.g., heat shock and f3-tubulin (3, 4)-and, conversely, can repress enhancer-stimulated transcription from the promoters of certain other genes-e.g., immunoglobulin and simian virus 40 early promoters (5-7). How these modes of positive and negative regulation by ElA may be involved in ElA-dependent viral transformation and tumorigenesis (reviewed in ref . 8)

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“…Most of the functions attributed to DBP are based on studies of viral mutants containing lesions in the E2A gene. These mutants can be grouped into four general classes (70). The first class consists of the temperature-sensitive (ts) mutants located in the C-t.…”

mentioning

confidence: 99%

Isolation and characterization of a viable adenovirus mutant defective in nuclear transport of the DNA-binding protein

Cleghon

Voelkerding

Morin

et al. 1989

J Virol

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The isolation and characterization of an adenovirus mutant, Ad5dl802rl, containing two independent deletions in the 72-kilodalton (kDa) DNA-binding protein (DBP) gene is described. The two deletions remove amino acids 23 through 105 of DBP, resulting in the production of a 50-kDa product. Expression of this truncated DBP was delayed 12 to 24 h compared with that of the 72-kDa protein produced by wild-type adenovirus type 5. The DBP was located primarily in the cytoplasm of infected cells, whereas the wild-type product was predominantly nuclear. Therefore, DBP appears to contain a nuclear localization signal within the deleted region. Ad5dl802rl DNA synthesis, viral late gene expression, and virus production were all delayed 12 to 24 h and were approximately 10-fold lower than with wild-type adenovirus type 5. These phenotypic properties c,an be accounted for by the delay in synthesis and the inefficient accumulation of the 50-kDa DBP within the nucleus of infected cells. The truncated DBP also lacks the majority of amino acids which are phosphorylated in the normal protein. The loss of these phosphorylation sites does not appear to seriously impair the ability of the protein to carry out its functions.

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Adenovirus Genetics

Cited by 18 publications

References 262 publications

Latent infection of KB cells with adeno-associated virus type 2

Latent infection of KB cells with adeno-associated virus type 2

The 289-amino acid E1A protein of adenovirus binds zinc in a region that is important for trans-activation.

Isolation and characterization of a viable adenovirus mutant defective in nuclear transport of the DNA-binding protein

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