Pfeifer S, Pettersson R F, Kallio A, Oker-Blom N, Vaheri A
J Virol. 1981 Nov;40(2):533-40. doi: 10.1128/JVI.40.2.533-540.1981.
We have analyzed the structure of OK10-BM virus, an avian acute leukemia virus produced by a bone marrow-derived cell line of macrophage origin, and compared it with that of OK10 AV, an associated virus originally present in the OK10 virus stock. The RNAs of OK10-BM virus and OK10 AV had the same mobility in agarose gels, corresponding to 8.0 to 8.5 kilobases, a size considerably larger than that of the transforming component (5 to 6 kb) of most other avian acute leukemia viruses. Fingerprint analysis showed a close relationship between OK10-BM virus and OK10 AV RNAs. The polypeptide compositions of OK10-BM and OK10 AV viruses were similar except for the envelope glycoproteins. In analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the large envelope glycoprotein of OK10-BM virus migrated at M(r) = 78,000 (gp78), whereas OK10 AV had the characteristic 85,000-dalton glycoprotein (gp85) of nondefective avian leukemia viruses. gp78 was weakly labeled with methionine, glycine, proline, or mannose, suggesting that purified OK10-BM virus had reduced amounts of the modified envelope glycoprotein. In cell-free rabbit reticulocyte lysates, OK10-BM virion RNA directed the synthesis of a 200,000-dalton polypeptide (p200), a 180,000-dalton polypeptide (pr180), and a 76,000-dalton polypeptide (pr76), whereas OK10 AV RNA gave rise only to pr180 and pr76, suggesting that p200 may represent an OK10-BM-encoded transforming protein. No biochemical evidence for the presence of an associated helper virus was found in the OK10-BM virus population produced by the macrophage cell line. However, when OK10-BM virus was serially passaged in chicken embryo fibroblasts, a virus having structural properties similar to those of OK10 AV (OK10 AV-specific oligonucleotides and gp85) appeared after three passages. Moreover, nonproducer clones of transformed cells could be readily obtained in OK10-BM virus-infected quail cell cultures. It is thus likely that the bone marrow-derived macrophage cell line produces a transforming virus defective in its env gene and low amounts of an associated helper virus, which upon transfer to fibroblasts is preferentially replicated.
我们分析了OK10 - BM病毒的结构,该病毒是一种由巨噬细胞来源的骨髓衍生细胞系产生的禽急性白血病病毒,并将其与OK10 AV的结构进行了比较,OK10 AV是最初存在于OK10病毒株中的一种相关病毒。OK10 - BM病毒和OK10 AV的RNA在琼脂糖凝胶中的迁移率相同,对应于8.0至8.5千碱基,这一大小明显大于大多数其他禽急性白血病病毒的转化成分(5至6 kb)。指纹分析表明OK10 - BM病毒和OK10 AV的RNA之间存在密切关系。除包膜糖蛋白外,OK10 - BM和OK10 AV病毒的多肽组成相似。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分析中,OK10 - BM病毒的大包膜糖蛋白在M(r)= 78,000处迁移(gp78),而OK10 AV具有非缺陷型禽白血病病毒特有的85,000道尔顿糖蛋白(gp85)。gp78用甲硫氨酸、甘氨酸、脯氨酸或甘露糖标记较弱,表明纯化的OK10 - BM病毒中修饰的包膜糖蛋白含量减少。在无细胞兔网织红细胞裂解物中,OK10 - BM病毒粒子RNA指导合成了一种200,000道尔顿的多肽(p200)、一种180,000道尔顿的多肽(pr180)和一种76,000道尔顿的多肽(pr76),而OK10 AV RNA仅产生pr180和pr76,这表明p200可能代表一种由OK10 - BM编码的转化蛋白。在巨噬细胞系产生的OK10 - BM病毒群体中未发现存在相关辅助病毒的生化证据。然而,当OK10 - BM病毒在鸡胚成纤维细胞中连续传代时,传代三次后出现了一种具有与OK10 AV相似结构特性(OK10 AV特异性寡核苷酸和gp85)的病毒。此外,在OK10 - BM病毒感染的鹌鹑细胞培养物中很容易获得转化细胞的非生产性克隆。因此,骨髓衍生的巨噬细胞系可能产生一种env基因有缺陷的转化病毒和少量相关辅助病毒,后者在转移到成纤维细胞后优先复制。