Expression of polyoma virus in heterokaryons between embryonal carcinoma cells and differentiated cells

Boccara, Martine; Kelly, Françoise

doi:10.1016/0042-6822(78)90342-2

Cited by 47 publications

(25 citation statements)

References 7 publications

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“…One block to productive infection of EC cells by wild-type polyoma virus occurs after virus adsorption and penetration but before viral early protein synthesis (4,5). Because with BstNI and fractionated in a 7.5% polyacrylamide gel.…”

Section: Discussionmentioning

confidence: 99%

Polyoma mutants that productively infect F9 embryonal carcinoma cells do not rescue wild-type polyoma in F9 cells

Fujimura¹,

Linney²

1982

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

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Section: Discussionmentioning

confidence: 99%

Polyoma mutants that productively infect F9 embryonal carcinoma cells do not rescue wild-type polyoma in F9 cells

Fujimura¹,

Linney²

1982

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

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“…They contain a tandem repeat (of 33 and 67 bp) of polyoma DNA sequences containing an A-T toGC bp point mutation (at nucleotide 5230) located at 69 map units in the genome. The Blangy mutants (11,12) that replicate in PCC4-aza-1 EC cells do not contain this A-T to G-C bp change (11) (1)(2)(3)(4)(5)(6)(7)(8)(9), suggesting that these viruses can be used to explore cellular control mechanisms operative during developmental changes.…”

mentioning

confidence: 99%

“…When pluripotent EC cells are permitted to differentiate, they produce infectious polyoma virus or can be transformed by SV40 and express early viral proteins (4)(5)(6)(7). The blockage to polyoma or SV40 gene expression in several different EC cell lines has been investigated (1)(2)(3)8).…”

mentioning

confidence: 99%

“…Although polyoma virus undergoes a productive infection and simian virus 40 (SV40) expresses its early proteins in most differentiated mouse cells, neither virus produces detectable levels of their early proteins in murine embryonal carcinoma cells (EC cells) (1)(2)(3). When pluripotent EC cells are permitted to differentiate, they produce infectious polyoma virus or can be transformed by SV40 and express early viral proteins (4)(5)(6)(7).…”

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confidence: 99%

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Isolation and characterization of polyoma host range mutants that replicate in nullipotential embryonal carcinoma cells.

Sekikawa¹,

Levine²

1981

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

128

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Polyoma wild-type virus replicates in most murine differentiated cells but fails to produce virus in murine embryonal carcinoma cells. Polyoma host range mutants have been isolated that replicate in several nullipotential embryonal carcinoma cell lines but fail to replicate in a pluripotential cell line. The final virus yield ofthese host range mutants is dependent upon the multiplicity of infection in both differentiated cels and nuirotent embryon carcinoma cells. Two independently derived host range mutants contain the same single base pair change (A T to G-C) and a 33-and 67-base pair duplication ofthose viral DNA sequences containing this point mutation. This duplication of viral DNA is located at 69 map units on the polyoma genome on the late gene side of the origin of viral DNA replication (70.5 map units). This type of mutation suggests several models to explain the polyoma host range restriction in embryonal carcinoma cells.Although polyoma virus undergoes a productive infection and simian virus 40 (SV40) expresses its early proteins in most differentiated mouse cells, neither virus produces detectable levels of their early proteins in murine embryonal carcinoma cells (EC cells) (1-3). When pluripotent EC cells are permitted to differentiate, they produce infectious polyoma virus or can be transformed by SV40 and express early viral proteins (4-7). The blockage to polyoma or SV40 gene expression in several different EC cell lines has been investigated (1-3, 8). In nullipotential F9 cells infected with SV40, low levels of viral transcriptional intermediates and RNA have been detected, and the SV40 RNA is not spliced into large and small tumor antigen (T antigen) m-RNAs (3). When F9 cells were stimulated to produce endoderm (9), then SV40 infection resulted in the production of spliced mRNAs, and the viral tumor antigens were synthesized (7). Polyoma infection of the pluripotent EC cell line PCC4-aza-1 (10) results in the production of very small amounts of viral RNA (11) compared to an infection of differentiated cells. Polyoma host range mutants have been isolated that produce infectious virus on PCC4-aza-1 cells (12) but fail to replicate on nullipotent F9 cells. This indicates that the block to polyoma early gene expression in two different EC cell lines, F9 and PCC4-aza-1, may be different.We report the isolation of polyoma host range mutants that were selected for growth on the nullipotential EC cell line F9. These mutants also replicate -on two other nullipotent EC cell lines, MH and FA, but do not produce virus in the pluripotent EC cell line PCC4-aza-1. The yield ofinfectious host range mutant viruses in both differentiated cells and EC cells was dependent upon the input multiplicity of infection (moi). Two independently derived host range mutants were shown, by restriction enzyme mapping, to contain an insertion in the polyoma genome between map coordinates 67.4 and 70.0. DNA sequence analysis in this region demonstrated that these insertions were 33-and 67-base pair (bp) duplications of...

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“…The mechanism by which EC cells block the viral cycle is not completely understood. Boccara and Kelly (2) have shown that heterocaryons between EC cells and differentiated mouse cells are fully susceptible to virus infections; it is therefore unlikely that viral restriction is due to a mechanism involving a diffusible repressor-type molecule. Furthermore, fusion of Py-infected EC cells with baby hamster kidney cells, which can express only Py early proteins, leads to virus production.…”

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confidence: 99%

T-antigen-independent replication of polyomavirus DNA in murine embryonal carcinoma cells.

Dandolo¹,

Aghion²,

Blangy³

1984

Mol. Cell. Biol.

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Expression of wild-type polyomavirus (Py) is restricted in murine embryonal carcinoma (EC) cells. The block appears to be located at the level of early transcription. Since no T antigen is produced, we investigated the fate of viral DNA upon infection of these cells; we showed that wild-type Py DNA replicates efficiently in all EC cells, probably via a T-antigen-independent mechanism. Furthermore, we studied, at permissive and restrictive temperatures, the replication of tsa (thermosensitive for T antigen) viral DNA of an in vitro-constructed deletion mutant lacking part of the early region coding sequences and of a double mutant carrying both the tsa mutation and the PyEC F9 mutation (allowing expression of early and late viral functions in EC cells does replicate in EC cells, although less efficiently than the DNA of PyEC mutants. This replication proceeds via a Tantigen-independent mechanism as confirmed by the behavior of a Py tsa mutant, which produces a thermosensitive large T antigen, of a deletion mutant which lacks part of the large T protein-coding sequence, and of a double mutant carrying both the tsa and the PyEC F9 mutations. We also show that the ability to replicate Py DNA in the absence of T antigen is lost when EC cells are allowed to differentiate. Finally, we discuss the involvement of both viral DNA sequences and cellular proteins in this phenomenon. MATERIALS AND METHODSCell cultures and virus strains. NIH 3T6 mouse cells and the EC cell lines PCC4-aza-1, F9, and LT1 (12,16,21) were grown in Dulbecco modified Eagle medium supplemented with 10% fetal calf serum (GIBCO Diagnostics, Madison, Wis.). F9 cells were grown on gelatin-coated (0.1%) dishes. All virus strains were propagated by infecting secondary mouse embryo cells at a multiplicity of 0.01 PFU per cell. The wild-type Py used in these studies was the large plaque strain A2 (10). The nucleotide numbers refer to the Py DNA sequence as reported by Soeda et al. (19). The PyEC mutant PyF9-1 has been previously described (13

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Expression of polyoma virus in heterokaryons between embryonal carcinoma cells and differentiated cells

Cited by 47 publications

References 7 publications

Polyoma mutants that productively infect F9 embryonal carcinoma cells do not rescue wild-type polyoma in F9 cells

Polyoma mutants that productively infect F9 embryonal carcinoma cells do not rescue wild-type polyoma in F9 cells

Isolation and characterization of polyoma host range mutants that replicate in nullipotential embryonal carcinoma cells.

T-antigen-independent replication of polyomavirus DNA in murine embryonal carcinoma cells.

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