Ability of a T-antigen transport-defective mutant of simian virus 40 to immortalize primary cells and to complement polyomavirus middle T in tumorigenesis.

1986 ◽  
Vol 59 (3) ◽  
pp. 655-659 ◽  
Author(s):  
J Vass-Marengo ◽  
A Ratiarson ◽  
C Asselin ◽  
M Bastin
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Primary Cells ◽  
Defective Mutant ◽  
Antigen Transport

Maintenance of cellular proliferation by adenovirus early region 1A in fibroblasts conditionally immortalized by using simian virus 40 large T antigen requires conserved region 1

1990 ◽  
Vol 10 (12) ◽  
pp. 6664-6673
Author(s):  
T E Riley ◽  
A Follin ◽  
N C Jones ◽  
P S Jat
Keyword(s):  
Cell Line ◽  
Growth Arrest ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Growth Defect ◽  
Primary Cells ◽  
Nonpermissive Temperature ◽  
Large T Antigen ◽  
Adenovirus E1a

Various mutants of adenovirus E1A were assayed for their ability to complement the growth defect at the nonpermissive temperature for the cell line tsa14 which was isolated by immortalizing rat embryo fibroblasts with the thermolabile large T antigen of tsA58. This cell line grows indefinitely at the permissive temperature but undergoes rapid growth arrest upon shift up to the nonpermissive temperature. Since this growth arrest can be overcome by introduction of wild-type simian virus 40 large T antigen, human papillomavirus 16 E7, and adenovirus E1A, the tsa14 cells provided an excellent system for defining regions of E1A necessary for complementation of the growth defect. We demonstrate that conserved region 1 (CR1) is the region of E1A required for complementation. While CR2 of E1A has been shown to be required for the immortalization of primary cells and is also necessary for the binding of the 105-kDa retinoblastoma protein, mutations within this region did not abrogate complementation of the growth defect. However, since both CR1 and CR2 have previously been shown to be absolutely required for immortalization of primary cells by adenovirus E1A, this evidence suggests that the tsa14 system assays for the maintenance of proliferation and that this requires CR1.

Quantitation of a 55K cellular protein: similar amount and instability in normal and malignant mouse cells

1982 ◽  
Vol 2 (7) ◽  
pp. 763-771
Author(s):  
P T Mora ◽  
K Chandrasekaran ◽  
J C Hoffman ◽  
V W McFarland
Keyword(s):  
Half Life ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Cellular Protein ◽  
T Antigen ◽  
Similar Amount ◽  
Parent Cell ◽  
Primary Cells ◽  
Quantitative Expression ◽  
Tumorigenic Cells

Quantitative expression of a specific 55,000 (55K)-molecular-weight cellular protein was studied in two groups of mouse embryo fibroblast (clonal) cells originating from two parent clones, one of which possessed high tumorigenicity and the other of which possessed very low tumorigenicity. From the clone with low tumorigenicity, tumor lines and clones were obtained by selecting rare spontaneously transformed highly tumorigenic (mutant) cells. Cells were labeled during exponential growth for 3 h at 37 degrees C, with [35S]methionine, and the cellular 55K protein was immunoprecipitated with a monoclonal antibody and quantitated. There were low and approximately equal amounts of 55K protein in cells (clones) with both low and high tumorigenicity from both groups of cells, and there was no correlation at all between quantitative expression of 55K protein and of cellular tumorigenicity. There was approximately 10- to 20-fold more 55K protein in all simian virus 40-transformed T antigen-positive derivative clones, as shown previously. The T antigen-negative revertant tumor lines and clones obtained by an immunological in vivo selection method had low amounts of 55K protein, similar to the parent cell before simian virus 40 transformation. In all of the T antigen-negative cells, including the highly tumorigenic cells, degradation (turnover?) of the 55K protein was rapid, and a half-life of 15 to 60 min was estimated from pulse-chase experiments. In all of the T antigen-positive cells the 55K protein was stable (half-life greater than 10 h). In primary cells established from the tumors induced by highly tumorigenic cells there was a very low or no detectable amount of the 55K protein. This is in contrast to the primary cells obtained from early murine embryos in which we have reported high amounts of (stable) 55K proteins.

scholarly journals Maintenance of cellular proliferation by adenovirus early region 1A in fibroblasts conditionally immortalized by using simian virus 40 large T antigen requires conserved region 1.

1990 ◽  
Vol 10 (12) ◽  
pp. 6664-6673 ◽  
Author(s):  
T E Riley ◽  
A Follin ◽  
N C Jones ◽  
P S Jat
Keyword(s):  
Cell Line ◽  
Growth Arrest ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Growth Defect ◽  
Primary Cells ◽  
Nonpermissive Temperature ◽  
Large T Antigen ◽  
Adenovirus E1a

Various mutants of adenovirus E1A were assayed for their ability to complement the growth defect at the nonpermissive temperature for the cell line tsa14 which was isolated by immortalizing rat embryo fibroblasts with the thermolabile large T antigen of tsA58. This cell line grows indefinitely at the permissive temperature but undergoes rapid growth arrest upon shift up to the nonpermissive temperature. Since this growth arrest can be overcome by introduction of wild-type simian virus 40 large T antigen, human papillomavirus 16 E7, and adenovirus E1A, the tsa14 cells provided an excellent system for defining regions of E1A necessary for complementation of the growth defect. We demonstrate that conserved region 1 (CR1) is the region of E1A required for complementation. While CR2 of E1A has been shown to be required for the immortalization of primary cells and is also necessary for the binding of the 105-kDa retinoblastoma protein, mutations within this region did not abrogate complementation of the growth defect. However, since both CR1 and CR2 have previously been shown to be absolutely required for immortalization of primary cells by adenovirus E1A, this evidence suggests that the tsa14 system assays for the maintenance of proliferation and that this requires CR1.

scholarly journals Establishment and characterization of transformed goat primary cells by expression of simian virus 40 large T antigen for orf virus propagations

PLoS ONE ◽  
2019 ◽  
Vol 14 (12) ◽  
pp. e0226105
Author(s):  
Yumiko Yamada ◽  
Guan-Ru Liao ◽  
Ching-Yu Tseng ◽  
Yeu-Yang Tseng ◽  
Wei-Li Hsu
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Orf Virus ◽  
Primary Cells ◽  
Large T Antigen

scholarly journals Archetype JC Virus Efficiently Replicates in COS-7 Cells, Simian Cells Constitutively Expressing Simian Virus 40 T Antigen

1998 ◽  
Vol 72 (7) ◽  
pp. 5335-5342 ◽  
Author(s):  
Kazuya Hara ◽  
Chie Sugimoto ◽  
Tadaichi Kitamura ◽  
Naoto Aoki ◽  
Fumiaki Taguchi ◽  
...  
Keyword(s):  
Large Scale ◽  
Jc Virus ◽  
Regulatory Region ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Sv40 T Antigen ◽  
Hemagglutination Assay ◽  
Viral Culture ◽  
Defective Mutant

ABSTRACT JC polyomavirus (JCV), the causative agent of progressive multifocal leukoencephalopathy (PML), is ubiquitous in humans, infecting children asymptomatically and then persisting in the kidney. Renal JCV is not latent but replicates to excrete progeny in the urine. The renal-urinary JCV DNAs carry the archetype regulatory region that generates various rearranged regulatory regions occurring in JCVs derived from the brains of PML patients. Tissue cultures that support the efficient growth of archetype JCV have not been reported. We studied whether archetype JCV could replicate in COS-7 cells, simian cells transformed with an origin-defective mutant of simian virus 40 (SV40). Efficient JCV replication, as detected by a hemagglutination assay, was observed in cultures transfected with five of the six archetype DNAs. The progeny JCVs could be passaged to fresh COS-7 cells. However, when the parental cells of COS-7 not expressing T antigen were transfected with archetype JCV DNAs, no viral replication was detected, indicating that SV40 T antigen is essential for the growth of JCV in COS-7 cells. The archetype regulatory region was conserved during viral growth in COS-7 cells, although a small proportion of JCV DNAs underwent rearrangements outside the regulatory region. We then attempted to recover archetype JCV from urine by viral culture in COS-7 cells. Efficient JCV production was observed in COS-7 cells infected with five of the six JCV-positive urine samples examined. Thus, COS-7 cells should be of use not only for the production of archetype JCV on a large scale but also for the isolation of archetype JCV from urine.

scholarly journals Two Independent Regions of Simian Virus 40 T Antigen Increase CBP/p300 Levels, Alter Patterns of Cellular Histone Acetylation, and Immortalize Primary Cells

2013 ◽  
Vol 87 (24) ◽  
pp. 13499-13509 ◽  
Author(s):  
M. T. Saenz Robles ◽  
C. Shivalila ◽  
J. Wano ◽  
S. Sorrells ◽  
A. Roos ◽  
...  
Keyword(s):  
Histone Acetylation ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Primary Cells

scholarly journals Quantitation of a 55K cellular protein: similar amount and instability in normal and malignant mouse cells.

1982 ◽  
Vol 2 (7) ◽  
pp. 763-771 ◽  
Author(s):  
P T Mora ◽  
K Chandrasekaran ◽  
J C Hoffman ◽  
V W McFarland
Keyword(s):  
Half Life ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Cellular Protein ◽  
T Antigen ◽  
Similar Amount ◽  
Parent Cell ◽  
Primary Cells ◽  
Quantitative Expression ◽  
Tumorigenic Cells

Quantitative expression of a specific 55,000 (55K)-molecular-weight cellular protein was studied in two groups of mouse embryo fibroblast (clonal) cells originating from two parent clones, one of which possessed high tumorigenicity and the other of which possessed very low tumorigenicity. From the clone with low tumorigenicity, tumor lines and clones were obtained by selecting rare spontaneously transformed highly tumorigenic (mutant) cells. Cells were labeled during exponential growth for 3 h at 37 degrees C, with [35S]methionine, and the cellular 55K protein was immunoprecipitated with a monoclonal antibody and quantitated. There were low and approximately equal amounts of 55K protein in cells (clones) with both low and high tumorigenicity from both groups of cells, and there was no correlation at all between quantitative expression of 55K protein and of cellular tumorigenicity. There was approximately 10- to 20-fold more 55K protein in all simian virus 40-transformed T antigen-positive derivative clones, as shown previously. The T antigen-negative revertant tumor lines and clones obtained by an immunological in vivo selection method had low amounts of 55K protein, similar to the parent cell before simian virus 40 transformation. In all of the T antigen-negative cells, including the highly tumorigenic cells, degradation (turnover?) of the 55K protein was rapid, and a half-life of 15 to 60 min was estimated from pulse-chase experiments. In all of the T antigen-positive cells the 55K protein was stable (half-life greater than 10 h). In primary cells established from the tumors induced by highly tumorigenic cells there was a very low or no detectable amount of the 55K protein. This is in contrast to the primary cells obtained from early murine embryos in which we have reported high amounts of (stable) 55K proteins.

Structure and synthesis of a simian virus 40 super T-antigen.

1982 ◽  
Vol 44 (3) ◽  
pp. 963-973 ◽  
Author(s):  
M Lovett ◽  
C E Clayton ◽  
D Murphy ◽  
P W Rigby ◽  
A E Smith ◽  
...  
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen
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