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. 1980 Apr;77(4):2274–2278. doi: 10.1073/pnas.77.4.2274

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

Kazuko Shiroki 1, Kaoru Segawa 1, Hiroto Shimojo 1
PMCID: PMC348696  PMID: 6246533

Abstract

A tumor (T) antigen, designated T antigen g, was visualized as fine fluorescent granules in nuclei of adenovirus type 12 (Ad12)-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 Ad12 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 Ad12-infected cells when tested with narrowly reacting T sera. Extracts of [35S]methioninelabeled infected cells were immunoprecipitated with H sera, and the resultant precipitate was analyzed by the two-dimensional gel electrophoresis technique of O'Farrell. The autoradiogram showed the presence of a cluster of several polypeptides (Mr 35,000-40,000, pI 5.0-5.5) that was absent in extracts of mock-infected cells. A similar autoradiogram of infected cells analyzed with narrowly reacting T sera showed the presence of a small polypeptide (Mr 10,000, pI 6.4), that was absent in extracts of mock-infected cells. The results show that Mr 35,000-40,000 polypeptides are components of T antigen g and a Mr 10,000 polypeptide is a component of T antigen f. Ad12-transformed cells showed a similar result. T antigen g was present and T antigen f was absent in HY cells. Both T antigen g and T antigen f were present in CY cells, which are rat cells completely transformed by the EcoRI-C endonuclease fragment (0-16 map units) of Ad12 DNA. The possible functions of these proteins are discussed.

Keywords: indirect immunofluorescence, immunoprecipitation, transforming gene product

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