Aviv H, Boime I, Leder P
Proc Natl Acad Sci U S A. 1971 Sep;68(9):2303-7. doi: 10.1073/pnas.68.9.2303.
Small amounts of encephalomyocarditis virus RNA direct a 50-fold increase in amino acid incorporation, in appropriately supplemented ascites tumor cell extracts, under conditions that give rise to authentic viral polypeptides. Incorporation in these crude extracts has a novel characteristic, namely, that it is almost entirely dependent upon the addition of exogenous tRNA. Further, this incorporation is restricted in that tRNA derived from ascites tumor cells or from rat liver permits translation of viral RNA, whereas tRNA from yeast or Escherichia coli does not. These translational barriers are due, at least in part, to an incompatibility between the tRNA of yeast and E. coli and the aminoacyl-tRNA synthetases of the ascites tumor cell. A more extensive basis for this incompatibility is suggested, however, by the failure of the E. coli aminoacyl-tRNA synthetases to restore viral RNA-directed protein synthesis in the presence of tRNA from E. coli, although the coli synthetases fully restore the poly(U)-directed synthesis of polyphenyl-alanine. The possible role that unique or favored codon classes might play in this restriction is considered, together with the implications of the observed requirement for tRNA.
在能产生真实病毒多肽的条件下,少量脑心肌炎病毒RNA可使适当添加补充物的腹水肿瘤细胞提取物中的氨基酸掺入量增加50倍。在这些粗提取物中的掺入具有一个新特点,即几乎完全依赖于添加外源tRNA。此外,这种掺入受到限制,因为源自腹水肿瘤细胞或大鼠肝脏的tRNA能使病毒RNA翻译,而来自酵母或大肠杆菌的tRNA则不能。这些翻译障碍至少部分是由于酵母和大肠杆菌的tRNA与腹水肿瘤细胞的氨酰tRNA合成酶不兼容所致。然而,尽管大肠杆菌的氨酰tRNA合成酶能完全恢复聚(U)指导的聚苯丙氨酸合成,但在存在来自大肠杆菌的tRNA时,其无法恢复病毒RNA指导的蛋白质合成,这提示了这种不兼容性更广泛的基础。文中考虑了独特或偏好密码子类别在这种限制中可能发挥的作用,以及观察到的对tRNA需求的影响。