Transformation of a rat cell line by an adenovirus 7 DNA fragment

Sekikawa, Kenji; Shiroki, Kazuko; Shimojo, Hiroto; Ojima, Setsuko; Fujinaga, Kei

doi:10.1016/0042-6822(78)90103-4

Cited by 42 publications

(21 citation statements)

References 15 publications

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“…Transformation of cells with purified restriction endonuclease fragments of virus DNA (Graham et al, 1974;van der Eb et al, 1977;Sekikawa et aL, 1978;Shiroki et aL, 1977) as well as studies on the effect of host-range and deletion mutants on the transforming activity of the virus (Graham et al, 1978) have located the virus genes responsible for cellular transformation to (approx.) the left-most 12 % of the virus genome.…”

Section: Introductionmentioning

confidence: 99%

Expression of Early Viral Gene Products in Adenovirus Type 12-infected and -transformed cells

Esche

Siegmann

1982

Journal of General Virology

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SUMMARYWe have analysed early viral gene products expressed in adenovirus type 12 (Adl2)-infected cells as well as in two Adl2-transformed hamster cell lines, and Adl2-induced rat tumour cell lines by cell-free translation of virus-specific RNA which was selected by hybridization to cloned restriction endonuclease fragments of virus DNA. Proteins synthesized in vitro were analysed by one-and twodimensional gel electrophoresis. It was found that RNA encoded by early region E1A directs the synthesis of at least eight polypeptides with apparent mol. wt. 38K, 36K, 30K, 28K, 26K, 25K, 24K and 22K. All these proteins are related to each other. E1B-specific RNA directs the synthesis of three proteins: 59K, 19K and 17K. Early region E2a codes for a 61K polypeptide which probably represents the single-strand DNA-binding protein of Adl2. RNA complementary to region E3 directs the synthesis of a 16K protein, and RNA transcribed from region E4 the synthesis of polypeptides with mol. wt. 20K, 18K and l l.5K. We have mapped a 67K polypeptide into the region within 11 to 28 map units (E2b). The analysis of proteins directed by virus-specific RNAs prepared from two Adl2-transformed hamster cell lines (T637, HA12/7) and one Adl2-induced rat tumour line (RBT12/3) showed that early regions E1 and E4 are expressed in all three Adl2-transformed cell lines. RNA transcribed from early regions E2 and E3 have been detected in lines T637 and RBT12/3. The virus RNA prepared from the Adl2-transformed cell lines directed synthesis of polypeptides with mol. wt. very similar to those of early virus proteins from infected cells. However, in all three Adl2-transformed cell lines mentioned above we have found RNAs which directed the synthesis of additional polypeptides of early regions E1 (34K) and E4 (25K, 24K) not detected in infected cells. The DNA sequence between 11 and 28 map units (coding for the 67K protein) is not expressed in the Ad 12-transformed cells.

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Section: Introductionmentioning

confidence: 99%

Expression of Early Viral Gene Products in Adenovirus Type 12-infected and -transformed cells

Esche

Siegmann

1982

Journal of General Virology

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“…Since then, T antigen has been used as a marker for Adl2-infected or transformed cells. Recently, evidence has accumulated showing that the transforming genes of human adenovirus are in only a small region of the left end in the viral genome (4)(5)(6)(7)(8)(9). T antigen is considered to be the product of the adenovirus transforming genes as analyzed in Ad2- (10)(11)(12), 14), or Adl2-infected cells (15,16).…”

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confidence: 99%

Two tumor antigens and their polypeptides in adenovirus type 12-infected and transformed cells

Shiroki¹,

Segawa²,

Shimojo³

1980

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

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A tumor (T) antigen, designated T antigen g, was visualized as fine fluorescent granules in nuclei of adenovirus type 12 (Adl2)infected cells by immunofluorescence with sera from rats bearing HY cell tumors (H sera). HY cells are rat cells incompletely transformed by the Acc I-H endonuclease fragment (0-4.7 map units) of Adl2 DNA. The antigen is different from the usually described T antigen, designated T antigen f, which is visualized as fluorescent flecks or filaments in both nucleus and cytoplasm of Adl-infected (17). With the background described above, we once again tried to detect antibody in sera from rats bearing HY cell tumors. We used indirect immunofluorescence, in which Adl2-infected cells were stained with rat sera, followed by dye-conjugated anti-rat IgG. After examination of many rat sera, we found that a small number of sera stained the antigen in Adl2-infected cells. However, the morphology of the antigen (tentatively named T antigen g) was different from that of T antigen (tentatively named T antigen f) reported until now (2, 3). It was fluorescent granules confined to the nucleus, in contrast to fluorescent flecks or filaments in the nucleus and cytoplasm. We concluded that the new T antigen must be different from the classic T antigen of Adl2.On the basis of this finding, we tried to identify T antigen polypeptides with the use of sera from hamsters bearing tumors or sera from rats bearing CY 2274The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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“…This deduction followed from our unpublished observations. In our system, the DNA of 0-3.6 map units of BAV3 genome was not necessary for transformation (7,16), whereas the same left-hand ends of human adenovirus genome were essential (20,23,28). We recently found that it is the left-hand end where r or 1 strand of each viral DNA is transcribed, possibly in the opposite direction, in BAV3 and human adenoviruses, respectively (manuscript in preparation), though the other mode of transcription of BAV3 DNA) sites, strands, and directions) was generally similar to human adenoviruses (18,21).…”

Section: Discussionmentioning

confidence: 98%

Biochemical Studies on Bovine Adenovirus Type 3. VI. Identification of Virus-Specific Early Proteins and Tumor Antigens

Niiyama

1981

Cell Struct. Funct.

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ABSTRACT. At least 13 species of bovine adenovirus type 3 (BAV3)-specific early proteins were detected when two dimensional electrophoregrams of the BAV3-infected and mock-infected cells were compared. The molecular weights and isoelectric points ranged from 15,000 to 80,000 and from 5.35 to 6.85, respectively. Using an immunoprecipitation method, analyses of T antigens were carried out using three types of anti T sera. These antisera were prepared from tumor-bearing mice as induced by cells transformed with whole or specific fragments of BAV3 DNA containing transforming gene(s), and characterized as being BAV3-specific by immunofluorescent studies. Proteins of an apparent molecular weight of 15,000 (15 K) and 54,000 (54 K) were identified as common immunoprecipitates between these antisera and extracts of the cells lytically infected with BAV3. Each of these proteins was found among BAV3-specific early proteins. These results suggest that the 15 K and 54 K proteins are BAV3 tumor antigens. On the other hand, in the BAV3-transformd cells, a protein with an apparent molecular weight of 80 K, instead of 15 K or 54 K protein, was detected, using the same method. Such observations in human adenoviruses have not been reported. The BAV3 15 K protein seems to be suitable for the study of physicochemical properties and functions of T antigen since it can be obtained in large amounts.

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Transformation of a rat cell line by an adenovirus 7 DNA fragment

Cited by 42 publications

References 15 publications

Expression of Early Viral Gene Products in Adenovirus Type 12-infected and -transformed cells

Expression of Early Viral Gene Products in Adenovirus Type 12-infected and -transformed cells

Two tumor antigens and their polypeptides in adenovirus type 12-infected and transformed cells

Biochemical Studies on Bovine Adenovirus Type 3. VI. Identification of Virus-Specific Early Proteins and Tumor Antigens

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