Schlegel R A, Iversen P, Rechsteiner M
Nucleic Acids Res. 1978 Oct;5(10):3715-29. doi: 10.1093/nar/5.10.3715.
Red cell-mediated microinjection has been used to study tRNA turnover in SV3T3 mouse cells and TC7 cells, an African green monkey kidney line. The turnover of endogenous tRNA, measured by labeling with 3H-methionine, was first-order with half-lives of approximately one day in SV3T3 and two days in TC7 cells. 32PtRNA isolated from E. coli or TC7 cells turned over at the same rate as endogenous tRNA when injected into either SV3T3 or TC7 cells. This demonstrates that cellular processes, not properties inherent to tRNAs, are responsible for the difference in tRNA turnover observed between SV3T3 and TC7 cells. These results further indicate that the mechanism of tRNA turnover in mammaliam cells does not distinguish prokaryotic from eukaryotic tRNAs. In contrast to unmodified tRNA, glyoxalated tRNA was rapidly degraded upon injection. Thus altered tRNA's, like altered proteins, are turned over more rapidly in animal cells.
红细胞介导的显微注射已被用于研究SV3T3小鼠细胞和非洲绿猴肾细胞系TC7细胞中的tRNA周转情况。通过用³H-甲硫氨酸标记来测量内源性tRNA的周转,其呈一级反应,在SV3T3细胞中的半衰期约为一天,在TC7细胞中为两天。从大肠杆菌或TC7细胞中分离出的³²P-tRNA,当注入SV3T3或TC7细胞时,其周转速度与内源性tRNA相同。这表明是细胞过程而非tRNA固有的特性导致了在SV3T3和TC7细胞中观察到的tRNA周转差异。这些结果进一步表明,哺乳动物细胞中tRNA周转的机制并不区分原核和真核tRNA。与未修饰的tRNA不同,经乙二醛化的tRNA在注射后会迅速降解。因此,与改变的蛋白质一样,改变的tRNA在动物细胞中的周转更快。