SV40 T-antigen is required for maintenance of immortal growth in SV40-transformed human fibroblasts.
Two lines of immortal human fibroblasts were isolated following transfection of TIG-3 cells with plasmid DNA, pMT-1ODtsA, that contained SV40 early gene with a deletion in replication origin and ts mutation in coding sequence for T-antigen. These cells continued proliferation at 34 degrees C, over 565 population doubling level (PDL) which is far over the limited division potential of untransformed normal TIG-3 of 70-80 PDL. When the culture temperature was shifted to 40 degrees C after 70 PDL, they ceased proliferation immediately. One of these immortal clones, SVts8, lost its ts phenotype after retransformation with wtT-antigen gene. These results indicated that the function of intact T-antigen is required for maintenance of immortal proliferation, at least in one of the SV40 transformed immortal clones.
Tsuyama
N
N
Department of Cellular and Molecular Biology, Hiroshima University School of Medicine, Japan.
Miura
M
M
Kitahira
M
M
Ishibashi
S
S
Ide
T
T
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000502:physiology*; D015139:Blotting, Southern; D002455:Cell Division; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D008858:Microscopy, Phase-Contrast; D009154:Mutation; D013539:Simian virus 40 / Q000235:genetics / Q000502:physiology*; D013696:Temperature; D014158:Transcription, Genetic; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming