Sompayrac L, Danna K J
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder.
Virology. 1991 Mar;181(1):412-5. doi: 10.1016/0042-6822(91)90516-e.
T147D is an SV40 mutant that encodes only the amino-terminal 147 amino acids of large T antigen and does not make small t antigen. We have constructed a recombinant retrovirus that expresses the T147D mutant protein. We show here that this virus can transform the established rat cell line, F111, in an agar assay with high efficiency. More importantly, we demonstrate that this retrovirus transforms secondary rat embryo fibroblasts to anchorage independence as efficiently as a recombinant retrovirus that expresses both wild-type large and small T antigens. These data indicate that in rat cells, the amino-terminal 147 amino acids of T antigen are sufficient for transformation. Further, since the T147D protein does not bind p53, we conclude that the association between T antigen and p53 is not required for the transformation of rat cells to anchorage-independent growth.
T147D是一种SV40突变体，它仅编码大T抗原的氨基末端147个氨基酸，不产生小t抗原。我们构建了一种表达T147D突变蛋白的重组逆转录病毒。我们在此表明，这种病毒在琼脂试验中能够高效转化已建立的大鼠细胞系F111。更重要的是，我们证明这种逆转录病毒将原代大鼠胚胎成纤维细胞转化为不依赖贴壁生长的效率，与表达野生型大T和小T抗原的重组逆转录病毒一样高。这些数据表明，在大鼠细胞中，T抗原的氨基末端147个氨基酸足以实现转化。此外，由于T147D蛋白不与p53结合，我们得出结论，大鼠细胞转化为不依赖贴壁生长并不需要T抗原与p53之间的结合。
