Isolation of polyomavirus mutants multiadapted to murine embryonal carcinoma cells

Melin, F; Pinon, H.; Kress, Chantal; Blangy, Daniel

doi:10.1128/jvi.53.3.862-866.1985

Cited by 17 publications

(9 citation statements)

References 21 publications

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“…C is the point mutation at nt 5204 present in the duplicated copy of PyEC-C201 and PyEC-C306. The sequence of the evlOOl strain differs from that of evlOOlh previously described by M,lin et al (27): the point mutation observed by Magnusson and Nilsson (22) at nt 5228 in the second copy of the evlOO1 B enhancer was not found. Therefore, evlOOl contains a perfect duplication of the wt PyV genome from nt 5100 to 48. mixture of both viruses, were subcultured and checked for T-antigen expression at intervals.…”

Section: Resultsmentioning

confidence: 55%

“…PyEC-M112 has a genome structure very similar to that of the previously described mutant PyEC-M206 (27) but is replicated and transcribed more efficiently in all EC cell lines. The difference between the two viruses is most striking in F9 cells.…”

Section: Discussionmentioning

confidence: 69%

“…Cell lines and viruses. PCC4, F9, LT1 EC cells, and 3T6 cells were cultured as previously described (27). The origin and differentiation potential of the EC cell lines are indicated in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…We have reported that a mutant isolated on PCC4 cells, PyEC-M206, had acquired a multiadapted phenotype and expressed T antigen in all EC cell lines tested, including the F9 cell line; still, it did not harbor the characteristic PyEC-F9 rearrangements (27). Although it displayed an extended host range with regard to early viral gene expression, this mutant was unable to produce a visible lytic effect in EC cells; this result indicated that additional or different mutations might be required to obtain viruses which would be as efficient in EC cells as wt PyV in differentiated mouse fibroblasts.…”

mentioning

confidence: 97%

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Host range specificity of polyomavirus EC mutants in mouse embryonal carcinoma and embryonal stem cells and preimplantation embryos

Melin

Kemler

Kress

et al. 1991

J Virol

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New polyomavirus mutants (PyEC-C) selected on LT1 cells and exhibiting a strong cytopathic effect in all embryonal carcinoma (EC) cell lines tested have been isolated. They were derived by a sequence duplication event from a new multiadapted mutant isolated in PCC4 cells. A quantitative analysis of viral DNA replication and transcription in 3T6 and EC cell lines was performed to compare PyEC-C mutants and PyEC mutants previously isolated on F9 or PCC4 cell lines. Analysis of the results indicated that PyEC-C mutants were more efficient in all EC cell lines tested than all other PyEC mutants; on the contrary, they were less adapted to 3T6 cells than wild-type polyomavirus. In both 3T6 and EC cells, uncoupling between early transcription and viral DNA replication was observed; different viruses were shown to replicate with the same efficiency, while their levels of early transcripts differed by two orders of magnitude. Attempts to correlate the genome structure of the mutants with their biological properties indicate that duplication of protein-binding sequences is not the only event responsible for their phenotype. PyEC mutants were also analyzed with respect to their interactions with early mouse embryos and embryonal stem (ES) cell lines derived from the inner cell mass of blastocysts. They showed different degrees of expression in ES cells and preimplantation embryos. ES cells were most efficiently infected and lysed by mutants which exhibit both a multiadapted and a lytic phenotype in EC cells. Preimplantation embryos were not permissive to any PyEC mutants. However, EC-multiadapted mutants were infectious in blastocysts after two days of in vitro culture.

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Section: Resultsmentioning

confidence: 55%

Section: Discussionmentioning

confidence: 69%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 97%

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Host range specificity of polyomavirus EC mutants in mouse embryonal carcinoma and embryonal stem cells and preimplantation embryos

Melin

Kemler

Kress

et al. 1991

J Virol

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“…Differentiation of EC cells leads to cell types that are permissive for polyoma infection (38)(39)(40)(41). Several host range mutants of polyoma that infect undifferentiated EC cells have been isolated and characterized (42)(43)(44)(45)(46)(47). All of these mutants, designated PyEC mutants, have DNA sequence alterations within the polyoma enhancer.…”

Section: Introductionmentioning

confidence: 99%

Nuclear activity from F9 embryonal carcinoma cells binding specifically to the enhancers of wild-type polyoma virus and PyEC mutant DNAs

Fujimura¹

1986

Nucl Acids Res

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Although wild-type polyoma virus does not productively infect murine embryonal carcinoma (EC) cells, a number of mutants (PyEC mutants) that do infect undifferentiated EC cells have been isolated. All PyEC mutants have DNA sequence alterations within the enhancer region of the viral genome. This report describes an activity present in nuclear extracts of F9 EC cells which, by "footprint" analyses, binds specifically to a small region of about 20 base pairs (nucleotides 5180-5200) within the subregion of the polyoma enhancer designated as the B or beta element. While no difference in binding of factor was detected between wild-type polyoma enhancer and the enhancers of the PyEC mutants, PyF111 and PyF441, which had been selected for productive infection of F9 cells, definite differences between wild-type and mutants were observed in the digestion patterns of their naked DNAs with either DNAase I or exonuclease III. This difference was restricted to the region around the point mutation (nucleotide 5258) common to these mutant DNAs.

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Common features of polyomavirus mutants selected on PCC4 embryonal carcinoma cells.

Melin¹,

Pinon²,

Reiss

et al. 1985

The EMBO Journal

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The genomic rearrangements of six polyomavirus mutants selected on PCC4 embryonal carcinoma cells have been compared and their common characteristics pointed out. All mutants show a duplication which includes at least the adenovirus type 5 (Ad5) ElA-like enhancer core sequence plus a deletion of variable size and location. The presence of the second enhancer core sequence, the SV40-like enhancer, is not required for expression of the PyEC PCC4 phenotype. Two of these mutants are also able to express polyomavirus T antigen on F9 and LT1 cells. Multiadaptation seems to require the duplication of the Ad5 ElA-like core sequence, the maintenance of the SV40-like core sequence and a local change in DNA stability.

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Isolation of polyomavirus mutants multiadapted to murine embryonal carcinoma cells

Cited by 17 publications

References 21 publications

Host range specificity of polyomavirus EC mutants in mouse embryonal carcinoma and embryonal stem cells and preimplantation embryos

Host range specificity of polyomavirus EC mutants in mouse embryonal carcinoma and embryonal stem cells and preimplantation embryos

Nuclear activity from F9 embryonal carcinoma cells binding specifically to the enhancers of wild-type polyoma virus and PyEC mutant DNAs

Common features of polyomavirus mutants selected on PCC4 embryonal carcinoma cells.

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