Herrera F, Adamson R H, Gallo R C
Proc Natl Acad Sci U S A. 1970 Dec;67(4):1943-50. doi: 10.1073/pnas.67.4.1943.
Uptake of tRNA (Escherichia coli) was demonstrated in the murine leukemia, L1210 and in a human lymphoblast (NC-37) cell lines. In both cell lines, uptake of tRNA was rapid, reaching a maximum within 45 sec, and was linear with concentration up to about 50 mug/ml. This uptake of exogenous tRNA apparently was not due to altered membrane permeability or impaired cell viability, nor to ribonuclease degradation of the macromolecule. Furthermore, about 20% of the tRNA taken up by the cells remains functional and apparently intact. This was demonstrated by: (a) acylation with E. coli aminoacyl-tRNA synthetases; (b) methylation with leukemic cell tRNA-methylases, and (c) demonstrating (14)C-labeled 4S RNA in the cytoplasmic fraction of the leukemic cells after the addition of E. coli [(14)C]tRNA. The results demonstrate that tRNA can enter mammalian cells and suggest that an energy independent, carrier-mediated, mechanism may be operative.
在小鼠白血病L1210细胞系和人淋巴母细胞(NC - 37)细胞系中均证实了(大肠杆菌)tRNA的摄取。在这两种细胞系中,tRNA的摄取迅速,在45秒内达到最大值,并且在浓度高达约50微克/毫升时呈线性关系。这种外源性tRNA的摄取显然不是由于膜通透性改变或细胞活力受损,也不是由于大分子的核糖核酸酶降解。此外,细胞摄取的tRNA中约20%仍保持功能且显然完整。这通过以下方式得到证明:(a)用大肠杆菌氨酰 - tRNA合成酶进行酰化;(b)用白血病细胞tRNA甲基化酶进行甲基化,以及(c)在添加大肠杆菌[¹⁴C]tRNA后,在白血病细胞的细胞质部分中检测到¹⁴C标记的4S RNA。结果表明tRNA可以进入哺乳动物细胞,并提示可能存在一种不依赖能量、由载体介导的机制。