Adenovirus core proteins

Jm, Weber; Khittoo, Govindranathsing; Ar, Bhatti

doi:10.1139/m83-039

Cited by 21 publications

(12 citation statements)

References 23 publications

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“…The adenovirus nucleoprotein core consists of double stranded genomic DNA, three highly basic viral proteins VII, V, and μ (mu), as well as protein IVa2 and the 55-kDa terminal protein (Amin et al, 1977; Brown et al, 1975; Hosokawa and Sung, 1976; Maizel et al, 1968; Prage and Pettersson, 1971; Rekosh et al, 1977; Russell et al, 1968; Weber et al, 1983; Zhang et al, 2001). Protein VII is the major protein component of the core, with an estimated 1,070 copies present per virion (Everitt et al , 1973).…”

Section: Introductionmentioning

confidence: 99%

Accurate single-day titration of adenovirus vectors based on equivalence of protein VII nuclear dots and infectious particles

Walkiewicz

Morral

Engel

2009

Journal of Virological Methods

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SummaryProtein VII is an abundant component of adenovirus particles and is tightly associated with the viral DNA. It enters the nucleus along with the infecting viral genome and remains bound throughout early phase. Protein VII can be visualized by immunofluorescent staining as discrete dots in the infected cell nucleus. Comparison between protein VII staining and expression of the 72 kDa DNA binding protein revealed a one-to-one correspondence between protein VII dots and infectious viral genomes. A similar relationship was observed for a helper-dependent adenovirus vector expressing green fluorescent protein. This relationship allowed accurate titration of adenovirus preparations, including wild-type and helper-dependent vectors, using a one-day immunofluorescence method. The method can be applied to any adenovirus vector and gives results equivalent to the standard plaque assay.

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Section: Introductionmentioning

confidence: 99%

Accurate single-day titration of adenovirus vectors based on equivalence of protein VII nuclear dots and infectious particles

Walkiewicz

Morral

Engel

2009

Journal of Virological Methods

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“…Adenoviruses do not contain histones. Viral DNA is condensed into a core structure within the virus particle by viral basic proteins V, VII and X (Vayda et al, 1983;Weber et al, 1983). Electron microscopy reveals a beaded, nucleosome-like core structure, but micrococcal nuclease (MN) digestion yields DNA fragments of monomer and heterogeneous lengths unlike the regular repeating pattern obtained with host cell chromatin (Mirza & Weber, 1982;Vayda et al, 1983).…”

Section: Introductionmentioning

confidence: 99%

The Structure of Adenovirus Chromatin in Infected Cells

Déry¹,

Tóth²,

Brown³

et al. 1985

Journal of General Virology

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SUMMARYThe structure of adenovirus chromatin in infected cells was studied by micrococcal nuclease digestion and hybridization with virus-specific probes. In the early phase of infection (5 h) a significant proportion of viral molecules was organized like actively transcribed cellular chromatin. As expected for a transcriptionally active population of molecules, even at high multiplicity of infection the nucleosomal repeating pattern was less distinct than in a transformed cell which contained the corresponding but less active genomic region. The observed repeating pattern in infected cells was unlikely to be due to integrated molecules since less than 0.07~ of input genomes became associated with cellular DNA. After the onset of viral DNA replication, the pool of viral chromatin organized like cellular chromatin rapidly increased. In addition, newly replicated molecules also maintained the cellular chromatin-like organization as measured by [3H]thymidine incorporation after the cessation of cellular DNA synthesis. These data suggest that newly replicated viral molecules are organized by histones into cell-like chromatin throughout the infection cycle. Coincident with the peak of viral DNA and core protein synthesis, and the decline of histone synthesis, the late, core-like non-repeating viral chromatin became dominant, increasingly obscuring the underlying repeating pattern. Experiments suggest that this late chromatin is destined for encapsidation, that the early chromatin persists and that viral core proteins do not displace histones on viral DNA. A model is proposed suggesting that transcription and type I replication occur on histone-condensed templates, while type II replication products late in infection are condensed by core proteins and are destined for encapsidation.

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“…Interestingly, disappearance of the dots required ongoing RNA synthesis but not DNA synthesis. Taken together these data indicate that protein VII has an ongoing role during early phase and the beginning of DNA replication.The adenovirus nucleoprotein core consists of doublestranded genomic DNA, the highly basic viral proteins VII, V, and (mu), as well as protein IVa2 and the 55-kDa terminal protein (1,5,19,25,31,33,34,42). Protein VII is the major protein component of the core with an estimated 1,070 copies present per virion (9) and is primarily responsible for establishing viral chromatin structure.…”

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confidence: 99%

Adenovirus Protein VII Functions throughout Early Phase and Interacts with Cellular Proteins SET and pp32

Xue

Johnson

Ornelles

et al. 2005

J Virol

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Adenovirus protein VII is the major component of the viral nucleoprotein core. It is a highly basic nonspecific DNA-binding protein that condenses viral DNA inside the capsid. We have investigated the fate and function of protein VII during infection. "Input" protein VII persisted in the nucleus throughout early phase and the beginning of DNA replication. Chromatin immunoprecipitation revealed that input protein VII remained associated with viral DNA during this period. Two cellular proteins, SET and pp32, also associated with viral DNA during early phase. They are components of two multiprotein complexes, the SET and INHAT complexes, implicated in chromatin-related activities. Protein VII associated with SET and pp32 in vitro and distinct domains of protein VII were responsible for binding to the two proteins. Interestingly, protein VII was found in novel nuclear dot structures as visualized by immunofluorescence. The dots likely represent individual infectious genomes in association with protein VII. They appeared within 30 min after infection and localized in the nucleus with a peak of intensity between 4 and 10 h postinfection. After this, their intensity decreased and they disappeared between 16 and 24 h postinfection. Interestingly, disappearance of the dots required ongoing RNA synthesis but not DNA synthesis. Taken together these data indicate that protein VII has an ongoing role during early phase and the beginning of DNA replication.The adenovirus nucleoprotein core consists of doublestranded genomic DNA, the highly basic viral proteins VII, V, and (mu), as well as protein IVa2 and the 55-kDa terminal protein (1,5,19,25,31,33,34,42). Protein VII is the major protein component of the core with an estimated 1,070 copies present per virion (9) and is primarily responsible for establishing viral chromatin structure. It can potently condense DNA in vitro and in vivo and also repress transcription (3,6,21,35). This is consistent with the highly condensed configuration of viral chromatin found within the virion. When delivered to the nucleus, the chromatin is silent prior to stimulation by the viral transcriptional activator E1A (13).We and others have reported that protein VII from infectious viral particles enters the nucleus along with viral DNA and remains associated with it, suggesting that the protein VII-DNA complex is the substrate for transcriptional activation by E1A during early phase (8,15,18,21). Moreover we found that protein VII can associate with E1A protein in vitro (21).In this report we have studied the fate and function of protein VII during the early phase of infection. We demonstrate that virus-derived "input" protein VII persists in the nucleus throughout early phase and the beginning of viral DNA replication, suggesting that it has an ongoing role in gene regulation and perhaps DNA replication. During this period protein VII is found in discrete nuclear dot structures and viral DNA continues to associate with protein VII.We have also investigated the association of protein VII with cel...

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Adenovirus core proteins

Cited by 21 publications

References 23 publications

Accurate single-day titration of adenovirus vectors based on equivalence of protein VII nuclear dots and infectious particles

Accurate single-day titration of adenovirus vectors based on equivalence of protein VII nuclear dots and infectious particles

The Structure of Adenovirus Chromatin in Infected Cells

Adenovirus Protein VII Functions throughout Early Phase and Interacts with Cellular Proteins SET and pp32

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