Analysis of the Events Leading to SV40-Induced Chromosome Replication and Mitosis in Primary Mouse Kidney Cell Cultures

May, Evelyne; May, Pierre; Weil, Roger

doi:10.1073/pnas.68.6.1208

Cited by 21 publications

(12 citation statements)

References 14 publications

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“…I t is known that T-antigen plays a crucial role for initiation of SV40 DNA synthesis (23) but is insufficient for replication of virM progeny DNA which also needs a cellular factor(s) (14,16,17,25). The ability of hybrid and superhybrid cell lines to synthesize SV40V-antigen after superinfeetion, indicates that such a factor(s) is expressed at least in a small proportion of cells.…”

Section: Diseussionmentioning

confidence: 99%

Expression of ?early? and ?late? viral functions in a somatic cell hybrid between a mouse cell and a spontaneous yielder SV 40-transformed Chinese hamster cell

Suárez

Lavialle

Estrade

et al. 1978

Archives of Virology

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A somatic cell hybrid (Cl. 6d) was originated from the fusion of mouse 3T3-4E) and spontaneous yielder SV 40-transformed Chinese hamster (CHK/SVLP AG) cells. During the early stages of its history, the C1. 6d hybrid underwent a rapid chromosome loss, preferentially loosing hamster chromosomes. This was not a constant tendency of the hybrid cells. As the parental CHK)SVLP AG cells, the hybrid cells were always found 100 per cent SV40 T-antigen positive. While CHK/SVLP AG cells infectious SV 40 DNA, V-antigen and virus were regularly detected, in the hybrid cells only infectious DNA was occasionally detected. This was not due either to the loww of an essential Chinese hamster gene(s) or to the presence of an inhibiting mouse cell component(s); it was apparently the consequence of inability of the cells to properly activate the resident SV 40 genome(s). After superinfection with SV 40 DNA, the hybrid cells-though capable of synthesizing SV 40 V-antigen--were unable to ensure virus assembly. Experimental evidence was obtained suggesting that SV 40 maturation is dependent of a cellular function(s).

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Section: Diseussionmentioning

confidence: 99%

Expression of ?early? and ?late? viral functions in a somatic cell hybrid between a mouse cell and a spontaneous yielder SV 40-transformed Chinese hamster cell

Suárez

Lavialle

Estrade

et al. 1978

Archives of Virology

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“…Detection of SV40-induced-tumour (T) and capsid antigens Cells grown on cover slips (2cm x 2cm) were subjected to immunofluorescence tests by using either anti-T or anti-capsid serum as described previously (May et al, 1971).…”

Section: Preparation Ofsubcultures and Clonal Derivativesmentioning

confidence: 99%

Establishment of rat pancreatic endocrine cell lines by infection with simian virus 40

Niesor¹,

Wollheim²,

Mintz³

et al. 1979

Biochemical Journal

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The feasibility of infection and transformation by SV40 (simian virus 40) of primary cell cultures derived from newborn-rat pancreas was investigated. As judged by the presence of intranuclear SV40 T-antigen, exposure to the virus resulted specifically in infection and transformation of epithelioid (predominantly endocrine) cells. The transformed cells were subcultured (more than 64 passages) and cloned. Culture medium and acid/ethanol extracts of the cells did not contain detectable amounts of immunoreactive insulin after the third subculture. However, inoculation of such SV40-transformed pancreatic cells into immunodeficient rats results in tumours in which insulin production was partially restored through the passage in vivo, since the tumour cells contained and synthesized small amounts of immunoreactive insulin which co-migrated with an insulin marker on gel chromatography. Interestingly, the transformed cells maintained under tissue-culture conditions produced a protein immunologically related to insulin, soluble in aqueous buffer but insoluble in acid/ethanol. This 30000-dalton protein is too large to be a translation product of the rat preproinsulin 9 S mRNA. SV40-transformed pancreatic cells might prove useful in the investigation of the factors controlling and maintaining insulin biosynthesis.

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“…Cytoplasmic RNA was extracted by the phenol-chloroform method and fractionated by oligo(dT)-cellulose chromatography (Rosenthal, 1976) in order to obtain poly(A) + RNA. 3H-labeUed SV40 RNA was detected by hybridization to denatured SV40 DNA (present in excess) fixed on membrane filters (0-2/~g DNA/filter), as described by May et al (1973). Results presented in Table 2 show that IdUrd enhanced significantly the specific radioactivity of SV40 mRNA in the pretreated cells.…”

mentioning

confidence: 87%

“…Virus RNA polymerase activity was estimated as described by Shani et al (1977) in the Sarkosyl supernatant isolated from either IdUrd-pretreated or untreated cells, 24 h after infection. Since transcriptional activity was expected to be very low it was determined by hybridization of in vitro synthesized RNA to SV40 DNA immobilized on minifilters (May et al, 1973). The activity was expressed as 32p-UMP ct/min incorporated into hybridizable RNA per mg protein of Sarkosyl supernatant.…”

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confidence: 99%

Enhancement of Transcription of the SV40 Genome in Mouse Embryo Cells by Pretreatment with 5-Iodo-2'-deoxyuridine

Suárez¹,

Lange

Cassingéna

1981

Journal of General Virology

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SUMMARYTreatment of mouse embryo (ME) cells with 5-iodo-2'-deoxyuridine (IdUrd) before infection with SV40 virus, enhances T-antigen (T-Ag) production as detected by immunofluorescence and complement fixation. Cellular DNA and RNA synthesis are inhibited in both SV40 and mock-infected cells after IdUrd treatment. The analogue pretreatment significantly increases the amount of radiolabelled nuclear and cytoplasmic SV40-specific RNA and the RNA polymerase activity of the viral transcriptional complexes of the Sarkosyl supernatants, suggesting that the enhancement of SV40 T-Ag production in infected pretreated ME cells results from an increased synthesis of early virus RNA.It has been reported previously (Su~trez et al., 1976(Su~trez et al., , 1977) that treatment of fully, as well as semi-permissive, cells with 5-iodo-2'-deoxyuridine (IdUrd) before treatment with SV40 virus or virus DNA, resulted in a marked increase of T-antigen (T-Ag) production and virus yield. No effect of the drug on the replication of the virus was found when the experiments were carried out with non-permissive (mouse or Syrian hamster embryo) cells (Suarez et al., 1976). Experiments performed to determine whether IdUrd affects adsorption, penetration or decapsidation of SV40 in permissive or semi-permissive cells indicated that the analogue acts between the arrival of the virus DNA in the nucleus and T-Ag synthesis. Moreover, the enhanced production of T-Ag persisted in permissive cells even if cytosine arabinoside (Ara-C) was immediately added after infection (Su/lrez et al., 1977). These results suggest that the halogenated pyrimidine acts at an early step of the SV40 cycle, perhaps at the level of early virus transcription or translation.Mouse primary or secondary cultures are non-permissive for SV40. Infection with the virus does not lead to synthesis of virus DNA or production of late virus polypeptides, but substantial amounts of SV40 T-Ag and the early SV40 RNA which encodes this protein(s) can usually be detected. This infection leads, within a few weeks, to the appearance of SV40-transformed cell colonies (Smith et aL, 1979). Mouse cells infected with SV40 are therefore particularly useful for investigating the effects of IdUrd pretreatment on the expression of SV40 early functions.Mouse embryo (ME) fibroblasts were prepared from 12-to 14-day-old embryos of Swiss albino mice. Cultures were routinely grown in Eagle's minimal essential medium (MEM, Eurobio, Paris, France) supplemented with 10% calf serum. Secondary ME cells suspended in growth medium supplemented with 10% calf serum, were seeded at 4 × 106 or 2 x l0 s cells/culture in 10 cm diam. plastic dishes, in the presence or absence of 100/zg/ml IdUrd respectively, and incubated at 37 °C. Four days later, the medium was removed from the cultures and after trypsinization, the number of cells was determined. It was virtually the same in treated and untreated cultures. The cells were then infected with SV40 virus at 50 p.f.u./cell. After 2 h adsorption at 37 °C, excess virus inoc...

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Analysis of the Events Leading to SV40-Induced Chromosome Replication and Mitosis in Primary Mouse Kidney Cell Cultures

Cited by 21 publications

References 14 publications

Expression of ?early? and ?late? viral functions in a somatic cell hybrid between a mouse cell and a spontaneous yielder SV 40-transformed Chinese hamster cell

Expression of ?early? and ?late? viral functions in a somatic cell hybrid between a mouse cell and a spontaneous yielder SV 40-transformed Chinese hamster cell

Establishment of rat pancreatic endocrine cell lines by infection with simian virus 40

Enhancement of Transcription of the SV40 Genome in Mouse Embryo Cells by Pretreatment with 5-Iodo-2'-deoxyuridine

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