Identification of two nuclear subclasses of the adenovirus type 5-encoded DNA-binding protein

Voelkerding, Karl V.; Klessig, Daniel F.

doi:10.1128/jvi.60.2.353-362.1986

Cited by 75 publications

(42 citation statements)

References 47 publications

(52 reference statements)

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“…Previous light and electron microscopic studies have established that the nuclei of adenovirus-infected cells contain several morphologically distinct inclusions (Martinez-Palomo et al, 1967;Sugawara et al, 1977;Moyne et al, 1978;Reich et al, 1983;Puvion-Dutilleul et al, 1984;Voelkerding and Klessig, 1986;Murti et al, 1990). Among these inclusions, particular attention has been focused on pleomorphic structures that contain DBP, a viral protein required for DNA replication (see Challberg and Kelly, 1989).…”

Section: Discussionmentioning

confidence: 97%

Adenovirus replication and transcription sites are spatially separated in the nucleus of infected cells.

et al. 1994

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We have visualized the intranuclear topography of adenovirus replication and transcription in infected HeLa cells. The results show that viral DNA replication occurs in multiple foci that are highly organized in the nucleoplasm. Pulse‐chase experiments indicate that newly synthesized viral double‐stranded DNA molecules are displaced from the replication foci and spread throughout the nucleoplasm, while the single‐stranded DNA replication intermediates accumulate in adjacent sites. Double‐labelling experiments and confocal microscopy show that replication occurs in foci localized at the periphery of the sites where single‐stranded DNA accumulates. The simultaneous visualization of viral replication and transcription reveals that the sites of transcription are predominantly separated from the sites of replication. Transcription is detected adjacent to the replication foci and extends around the sites of single‐stranded DNA accumulation. These data indicate that newly synthesized double‐stranded DNA molecules are displaced from the replication foci and spread in the surrounding nucleoplasm, where they are used as templates for transcription. Splicing snRNPs are shown to co‐localize with the sites of transcription and to be excluded from the sites of replication. This provides evidence that splicing of viral RNAs occurs co‐transcriptionally and that the sites of viral DNA replication are spatially distinct from the sites of RNA transcription and processing.

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Section: Discussionmentioning

confidence: 97%

Adenovirus replication and transcription sites are spatially separated in the nucleus of infected cells.

et al. 1994

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“…It has been reported that snRNP is present in viral centers containing ssDNA and RNA (25) and in viral replication centers (63). Since the 72K protein is present in these locations (9,45,62), we wished to investigate the possibility that the distribution pattern of snRNP relative to the 72K protein changes during the course of infection. We have observed snRNP in foci that also contain the 72K protein in a small fraction of cells in cultures at 18 h p.i.…”

Section: Distribution Of Non-snrnp Splicing Factors In Ad-infected Cementioning

confidence: 99%

Dynamic organization of splicing factors in adenovirus-infected cells

Bridge

Xia

Carmo‐Fonseca

et al. 1995

J Virol

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Adenovirus infection affects the nuclear distribution of host splicing factors. Late phase-infected cells contain discrete clusters of small nuclear ribonucleoproteins (snRNPs) that are separate from centers containing the viral 72-kilodalton DNA-binding protein (72K protein). In the present study, we demonstrate that these snRNP clusters also contain splicing factors from the SR protein family. We show that a previously described monoclonal antibody, 3C5, detects SR proteins. Furthermore, we demonstrate that late region 3 transcription occurs at a maximal rate in infected cultures in which greater than 90% of the cells contain the snRNP clusters, indicating that such cells are actively transcribing their late genes. During the onset of the late phase, the intranuclear distribution of splicing factors is very different from that seen after the late phase is established. When late viral transcription commences, cells with snRNP clusters are less prevalent than in cultures that are maintaining maximum levels of late transcription. Instead, a cell type which shows snRNPs, concentrated in foci that also contain the viral 72K DNA-binding protein is detected. This cell type disappears from cultures by 18 to 20 h after a high-multiplicity infection. These results suggest a dynamic organization of splicing factors in infected cells that can be correlated to the status of viral gene expression. Our work also provides an explanation for the differing results that have been published concerning the organization of splicing factors in the adenovirus-infected cell nucleus (L. F. Jiménez-García and D. L. Spector, Cell 73:47-59, 1993). During the present study we observed that a monoclonal antibody against the SC-35 protein, which was used by Jiménez-García and Spector to study the localization of the SC-35 splicing factor in adenovirusinfected cells, cross-reacts with the adenovirus 72K DNA-binding protein and is thus unsuitable for this type of study.

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“…Since the adenovirus DBP has been shown to colocalize with the viral DNA in replication factories (19), we used this as an independent means to probe for the localization of replication factories. HeLa cells were grown directly on microscope slides, infected with Ad2, fixed 24 hours post infection with methanol:acetone, and probed in situ using antiserum to Ad DBP followed by development with FITC-conjugated anti-Ig serum.…”

Section: Intranuclear Localization Of Ad2 Dna In Hela Cellsmentioning

confidence: 99%

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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Identification of two nuclear subclasses of the adenovirus type 5-encoded DNA-binding protein

Cited by 75 publications

References 47 publications

Adenovirus replication and transcription sites are spatially separated in the nucleus of infected cells.

Adenovirus replication and transcription sites are spatially separated in the nucleus of infected cells.

Dynamic organization of splicing factors in adenovirus-infected cells

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

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