Simian virus 40 large T-antigen function is required for induction of tetraploid DNA content during lytic infection

Friedrich, Thomas; Laffin, Judith; Lehman, John M.

doi:10.1128/jvi.66.7.4576-4579.1992

Cited by 29 publications

(17 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4), the cell population could move past the block and through the cell cycle as a synchronous cohort. Consistent with the idea that Tag can cause a transient cell cycle block, Tag expression has been shown to prevent mitosis during productive SV40 infection of monkey cells (5,24). In addition, we previously showed that Tag expression lengthens G 2 ϩ M cell cycle phase duration (28).…”

Section: Discussionsupporting

confidence: 80%

See 1 more Smart Citation

Cell cycle analysis of retroviral vector gene expression during early infection

Sladek¹,

Jacobberger²

1998

Cytometry

View full text Add to dashboard Cite

To examine the pattern of retroviral vector gene expression during early stages of infection, a Moloney murine leukemia virus (M‐MuLV)‐based vector that transcribes the simian virus 40 (SV40) large T antigen (Tag) gene from the viral long terminal repeat (LTR) was used to infect proliferating rodent fibroblasts. At various times after infection, cells were fixed and stained for Tag by indirect immunofluorescence, and for DNA using propidium iodide. Tag immunofluorescence and DNA content were both quantified by flow cytometry. The results showed that Tag expression was first detected exclusively in the late G1 and early S phases of the cell cycle, approximately 12 h postinfection. The infection was synchronous in that Tag‐expressing cells detected at 12 h, in late G1 and early S, moved as a discrete population through S phase, into the G2 + M phases of the cell cycle and then back into G1 during the next 8–10 h. The presence of a synchronous Tag‐expressing cell population suggests that, at time of infection, cells in certain phases of the cell cycle were more susceptible to infection than cells in other phases. This may be related to synchronizing events that must occur before viral genes are expressed in infected cells; one such event may be integration of viral DNA into cellular chromosomes (i.e., provirus formation) that requires cells to pass through an M phase. Cytometry 31:235–241, 1998. © 1998 Wiley‐Liss, Inc.

show abstract

Section: Discussionsupporting

confidence: 80%

“…Flow cytometric detection of viral gene expression in infected cells has been used to quantify virus number (4,8,21,30), to examine viral gene expression during the course of infection (4,14), and to evaluate the cellular phenotypes produced by expression of viral genes (5).…”

mentioning

confidence: 99%

Cell cycle analysis of retroviral vector gene expression during early infection

Sladek¹,

Jacobberger²

1998

Cytometry

View full text Add to dashboard Cite

show abstract

“…As a result, the number of surface-exposed large T molecules varied among different SV40-transformed cell lines derived from different species (Table 1 ). In the case of the murine cell line C57SV, the number of large T molecules per cell depended remarkably on the density of the culture used to prepare the cells, most probably reflecting the influence of the cell cycle on cell surface large T molecule frequency (6,16,19,23). Correlation of virus-specific fluorescence signals with cell-cycle phases has been previously shown for methanol-fixed murine SV40-infected CV-1 cells (6,19).…”

Section: Resultsmentioning

confidence: 74%

Quantitative cytofluorimetric determination of cell membrane‐associated large tumor antigen on SV40‐transformed cells

et al. 1995

View full text Add to dashboard Cite

The aim of this study was to quantitate the number of cell membrane-located SV40 large tumor antigen (large T) molecules of SV40-transformed cell lines by cytofluorimetric analysis. Five different SV40-transformed cell lines were labelled by either a biotin-or a fluorescein-conjugated monoclonal antibody, PAb1605, which is specmc for the large T carboxyterminus. The conjugated-antibody fluorescence signals of the stained large T molecules of transformed cells were measured via cytofluorimetry. Comparison of the fluorescence signals of calibrated beads bearing a known number of fluorescein molecules to the signals of conjugated PAb1605 antibodies bound on microbeads to a detbed number of IgG binding sites made it possible to determine the number of antibody-accessible large T molecules per SV40-transformed cell. The numbers ( X lo-*) found per cell were 1.0 (ELONA, hamster), 3.0 (VLM, mouse), 3.5 (mKSA, mouse), 11 (C57SV, mouse), and 5.5 (SVSO, human), respectively. Thus, the technique described allows a precise quantitation of surfaceexposed, antibody-accessible viral antigen expresdoll. 0 1995 WileyLiss, Inc.

show abstract

“…To determine the number of cells expressing T antigen, the cells were grown on coverslips, fixed with 70% acetone and 30% methanol at -20"C, and stained by indirect immunofluorescence with a polyclonal antibody described by Silver et al (18). The CV-1 cells, SV40 virus, and the infection procedure have been described previously (6,7,15).…”

Section: Methodsmentioning

confidence: 99%

“…When DNA content analyses were performed with flow cytometry, both permissive and nonpermissive cells are induced into multiple rounds of DNA synthesis without mitosis which produce a cell with a >G, DNA content (tetraploid S and tetraploid G,/M) (11,(14)(15)(16). Further, a temperature-sensitive mutant of SV40, tsA 30, was unable to initiate a second round of DNA synthesis a t the nonpermissive temperature (405°C) identifying a T antigen function necessary to bypass G,/M controls and initiate a second round of DNA synthesis (6).…”

mentioning

confidence: 99%

DNA content distribution of mouse cells following infection with polyoma virus

Lehman¹,

Laffin²,

Friedrich³

1994

Cytometry

View full text Add to dashboard Cite

Infection of primary to tertiary mouse embryo fibroblasts or mouse kidney cells with polyoma virus leads to stimulation of cellular DNA synthesis. When either confluent or growing mouse cells were infected, the monolayer cells were found to accumulate cells with a DNA content of S and G,/M phases of the cell cycle as assayed by flow cytometry. A similar pattern of DNA content was also observed in cells in the supernatant, which are probably cells replicating virus and dying. When compared with control cells, the infected monolayer and supernatant cells exhibited a population (5-27%) with a >G, DNA content. The increase in DNA content of these >G, cells was calculated to be an average of 26.7%, which is probably due to viral DNA. Polyoma contrasts with another papovavirus, SV40, which stimulates cells into DNA synthesis, with the majority of cells attaining a >G,/tetraploid DNA content, suggesting that there are differences in polyploidization between these two viruses.0 1994 Wiley-Liss, Inc.

show abstract

Simian virus 40 large T-antigen function is required for induction of tetraploid DNA content during lytic infection

Cited by 29 publications

References 30 publications

Cell cycle analysis of retroviral vector gene expression during early infection

Cell cycle analysis of retroviral vector gene expression during early infection

Quantitative cytofluorimetric determination of cell membrane‐associated large tumor antigen on SV40‐transformed cells

DNA content distribution of mouse cells following infection with polyoma virus

Contact Info

Product

Resources

About