Acquisition of enzyme function by mouse kidney cells abortively infected with papovavirus SV40.

Kit, Saul; Dubbs, D. R.; Piekarski, L; Torres, Ramón A.; Melnick, Joseph L.

doi:10.1073/pnas.56.2.463

Cited by 40 publications

(12 citation statements)

References 12 publications

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“…The induction of thymidine kinase coincided with the appearance of T antigen at both 37 ~ and 29~ this indicates that not only T antigen synthesis but also the induction of thymidine kinase are among the viral functions which are less temperature-dependent than the synthesis of V antigen. The observation that the thymidine kinase was induced at the decreased temperature at which none or very little infectious virus was formed complements the previous findings by KIT et al (3,12) and CARP et al (13) indicating that infectious virus production is not required for the induction of the enzyme.…”

Section: Effect Of Temperature On Synthesis Of T and V Antigens Of Pasupporting

confidence: 80%

Effect of temperature on synthesis of T and V antigens of papovavirus SV40 and induction of thymidine kinase

Vonka¹,

Kára²,

Kutinovà³

et al. 1969

Archiv f Virusforschung

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The syntheses of tumor (T) and viral capsid (V) antigens of papovavirus SV40 proceed differently at different temperatures. Using complement fixation (CF), I~T~A and 35EL~ICX (1) found that the peak T antigen titers were the same at 23 ~ as at 37~ although the first appearance of the T antigen was significantly delayed at the lower temperature. At 23~ the V antigen synthesis was markedly suppressed and the period between the appearance of detectable levels of the T and the V antigens was prolonged. The thermal separation of synthesis of T and V antigens was then applied to adenoviruses (2).Infection of monkey kidney cells with SV40 results in early synthesis not only of T antigen, but also of thymidine kinase. Their synthesis is not inhibited by drugs that prevent DNA synthesis (3, 4). However, KIT et al. (5) clearly demonstrated the non-identity of T antigen and thymidine kinase.In an attempt to separate the synthesis of T antigen and thymidine kinase, we studied their development in green monkey kidney (GMK) cells infected with SV40 and incubated at subnormal temperature. About 2 • 10 ~ GM_K cells in 5 ml of Parker's medium 199 supplemented with 2~o calf serum were seeded in 60 mm plastic Petri dishes, which were then incubated at 37~ in an atmosphere with 5% CO S. Some of the Petri dishes contained coverslips. Twenty-four hours after confluent cell sheets had been formed, the cultures were infected with SV40 (Baylor strain) at an input multiplicity of about 10 TCDs0 per cell. After 2 hours incubation

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Section: Effect Of Temperature On Synthesis Of T and V Antigens Of Pasupporting

confidence: 80%

Effect of temperature on synthesis of T and V antigens of papovavirus SV40 and induction of thymidine kinase

Vonka¹,

Kára²,

Kutinovà³

et al. 1969

Archiv f Virusforschung

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“…Primary cultures of mouse kidney and mouse embryo were prepared as previously described (Kit et a/., 1966~).…”

Section: Cell Linesmentioning

confidence: 99%

Transplantable mouse tumor line induced by injection of SV40‐transformed mouse kidney cells

Kit

Kurimura

Dubbs

1969

Intl Journal of Cancer

166

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A transplantable tumor of inbred mice was obtained by inoculating BALB/c mice subcutaneously with SV40‐transformed mouse kidney (mKS‐A) cells. Tumors were produced by mKS‐A cells in the 71st cell culture passage, but not by cells in the 26th passage. The tumor line has been serially passed in BALB/c mice 14 times. In vitro cell culture lines were derived from tumors after 1, 2, 8, 10 and 12 passages in mice. The tumors, as well as the In vitro tumor cell lines, contained SV40 T‐antigen, and sera from the tumor‐bearing mice contained antibodies to the SV40 T‐antigen. SV40 was rescued from the In vitro tumor cell lines after fusion with green monkey kidney (CV‐1) cells in the presence of UV‐irradiated Sendai virus. The In vitro tumor cell lines derived from mouse passages 8, 10 and 12 were used as SV40 virus; 2) SV40‐transformed cell lines; 3) primary mouse (BALB/c or Yale Swiss) kidney cells, or 4) primary mouse (BALB/c or Yale Swiss) embryo cells. These results showed that the tumor line and the In vitro tumor cell lines have the transplantation antigen.

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“…Infection of confluent, "contact-inhibited" primary mouse kidney (MK) cell cultures with simian virus 40 (SV40) induces the replication of cellular DNA, while little, if any, viral DNA, capsid protein, or progeny virus is produced (1,2). This abortive infection is followed, within a few weeks, by the appearance of foci of "transformed" cells (3,4).…”

mentioning

confidence: 99%

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

May

Weil

1971

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

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Abortive infection with simian virus 40 in confluent, "contact-inhibited", mouse kidney cell cultures was studied. The sequential events, in individual cells, are tentatively represented by the simplified scheme: Infection of confluent, "contact-inhibited" primary mouse kidney (MK) cell cultures with simian virus 40 (SV40) induces the replication of cellular DNA, while little, if any, viral DNA, capsid protein, or progeny virus is produced (1, 2). This abortive infection is followed, within a few weeks, by the appearance of foci of "transformed" cells (3,4).In this paper, evidence is presented that indicates that the abortive infection with SV40 actually triggers chromosome replication and mitosis. These results, and those presented elsewhere (5, and manuscript in preparation), show that this experimental system not only allows a detailed investigation of the early functions of SV40 and of their relation to the process of virus-induced cell "transformation", but that it is a promising model for the study of genetic regulation in mammalian cells. MATERIAL AND METHODSPrimary MK (6) cultures prepared from kidneys of 10-dayold white Swiss mice were grown in plastic Petri dishes (30-mm diameter, unless indicated otherwise) on glass coverslips (22 X 22 mm) (7). SV40 virus (large plaque, SVi strain) (8) and CV-1 cultures (an established monkey-kidney cell line) (9) were obtained in May 1969 from Dr. R. Cassingena. The virus used in our studies was grown and assayed for plaque-forming titer (PFU) on CV-1 cultures.Cultures were infected with SV40 virus by adding 0.15 ml of viral lysate and permitting adsorption to continue for 1 hr at 370C in a COrincubator. Unless indicated otherwise, the cultures were then covered with serum-free, reinforced Eagle's and Institut de Recherches medium (7) prewarmed to 370C. Essentially identical results were obtained when the cultures were washed after adsorption. The time at which virus was added to the cultures was considered to be time zero. Serial dilutions of the viral lysates were prepared in culture medium containing 2% calf serum (Microbiological Associates). In all experiments, control cultures were mock-infected and were treated subsequently in the same way as were the SV40-infected cultures.Immunofluorescence tests for SV40-specific T-antigen were performed according to the method of Fluorescein-conjugated anti-hamster globulin (from rabbits) was obtained from Progressive Laboratories, Inc., Baltimore, Md.Immunofluorescence tests for viral capsid antigen were performed as was described (7). The antiserum was obtained by repeated injection of highly purified SV40 capsids into the footpad of rabbits. The fluorescein-conjugated anti-rabbit globulin (from goats) was obtained from the Institut Pasteur (Paris). The relative number of cells with nuclear fluorescence for T-or capsid-antigen was determined in a Zeiss Ultraphot microscope. To obtain a value for each time interval a random sample of at least 1500 cells was counted.

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Acquisition of enzyme function by mouse kidney cells abortively infected with papovavirus SV40.

Cited by 40 publications

References 12 publications

Effect of temperature on synthesis of T and V antigens of papovavirus SV40 and induction of thymidine kinase

Effect of temperature on synthesis of T and V antigens of papovavirus SV40 and induction of thymidine kinase

Transplantable mouse tumor line induced by injection of SV40‐transformed mouse kidney cells

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

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