Beemon K, Hunter T
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3302-6. doi: 10.1073/pnas.74.8.3302.
In vitro translation of Rous sarcoma virus (RSV) virion RNA in the messenger-dependent reticulocyte lysate system yielded polypeptides that were not synthesized by translation of RNA from a transformation-defective deletion mutant of RSV. These RSV-specific products migrated on sodium dodecyl sulfate/polyacrylamide gels as two doublets of approximately 25,000 and 17,000 daltons. Synthesis of these proteins was not sensitive to inhibition by m7GTP; however, synthesis of the 76,000-dalton precursor of the internal structural proteins was sensitive to inhibition by m7GTP. Tryptic peptide maps showed the 25,000- and 17,000-dalton proteins to be related to one another but to be distinct from the 76,000-dalton protein. The 25,000-dalton protein was translated only from a polyadenylylated RNA of approximately 2500 nucleotides, whereas the 76,000-dalton protein was translated from 38S RNA, corresponding to the entire viral genome. A 180,000-dalton protein was also synthesized from 38S RSV virion RNA. From the absence of the 25,000- and 17,000-dalton proteins in the translation products of transformation-defective RSV RNA and the size of their RNA templates, we conclude that these proteins may be derived from coding sequences within the RSV src gene.
在信使依赖的网织红细胞裂解物系统中对劳氏肉瘤病毒(RSV)病毒粒子RNA进行体外翻译,产生了一些多肽,而这些多肽在RSV的转化缺陷型缺失突变体的RNA翻译过程中并未合成。这些RSV特异性产物在十二烷基硫酸钠/聚丙烯酰胺凝胶上迁移时呈现为两个双峰,分子量约为25,000道尔顿和17,000道尔顿。这些蛋白质的合成对m7GTP的抑制不敏感;然而,内部结构蛋白的76,000道尔顿前体的合成对m7GTP的抑制敏感。胰蛋白酶肽图谱显示,25,000道尔顿和17,000道尔顿的蛋白质彼此相关,但与76,000道尔顿的蛋白质不同。25,000道尔顿的蛋白质仅由约2500个核苷酸的多聚腺苷酸化RNA翻译而来,而76,000道尔顿的蛋白质则由38S RNA翻译而来,38S RNA对应整个病毒基因组。还从38S RSV病毒粒子RNA合成了一种180,000道尔顿的蛋白质。根据转化缺陷型RSV RNA翻译产物中不存在25,000道尔顿和17,000道尔顿的蛋白质以及它们RNA模板的大小,我们得出结论,这些蛋白质可能源自RSV src基因内的编码序列。