PRODUCT OF INVITRO TRANSLATION OF THE ROUS-SARCOMA VIRUS SRC GENE HAS PROTEIN-KINASE ACTIVITY
PRODUCT OF INVITRO TRANSLATION OF THE ROUS-SARCOMA VIRUS SRC GENE HAS PROTEIN-KINASE ACTIVITY
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DOI：
10.1128/jvi.30.1.311-318.1979
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发表时间：
1979-01-01
影响因子：
5.4
通讯作者：
BEEMON, K
中科院分区：
文献类型：
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作者：
SEFTON, BM;HUNTER, T;BEEMON, K
劳斯肉瘤病毒病毒粒子RNA的体外翻译导致蛋白激酶的合成，当与抗肿瘤血清免疫沉淀时，磷酸化免疫球蛋白H链。尽管病毒体RNA的体外翻译导致了许多被抗肿瘤血清识别的多肽的合成，但对照实验表明，只有当合成了src基因的多肽产物p60 src时，才发现免疫沉淀蛋白激酶活性。因此，具有类似性质的蛋白激酶与在网织红细胞裂解物中体外合成的p60src密切相关，就像它与从转化的鸡和哺乳动物细胞中获得的p60src一样。因此，这种关联极不可能是人为的。ts NY68是劳斯肉瘤病毒的突变体，其能够在36 ℃转化细胞。但不是在41 °。C，在30 ℃下ts NY68病毒体RNA的体外翻译。C导致免疫可沉淀p60src的有效合成，但免疫可沉淀蛋白激酶的合成非常低效。通过体外翻译野生型病毒体RNA获得的p60 src作为蛋白激酶的活性比从ts Ny68 RNA获得的活性高20倍以上。在ts Ny68的情况下，蛋白激酶活性缺乏与在高温下不能转化细胞的相关性表明，与p60 src相关的蛋白激酶活性确实对细胞转化至关重要。
In vitro translation of Rous sarcoma virus virion RNA resulted in the synthesis of a protein kinase which, when immunoprecipitated with antitumor serum, phosphorylated the immunoglobulin H chain. Even though in vitro translation of virion RNA resulted in the synthesis of a number of polypeptides which were recognized by antitumor serum, control experiments demonstrated that an immunoprecipitable protein kinase activity was found only when an immunoprecipitable p60src, the polypeptide product of the src gene, was synthesized. A protein kinase with similar properties was therefore intimately associated with p60src which was synthesized in vitro in the reticulocyte lysate, just as it is with p60src which is obtained from transformed chick and mammalian cells. It is therefore highly unlikely that this association is artifactual. ts NY68 is a mutant of Rous sarcoma virus which is able to transform cells at 36.degree. but not at 41.degree. C, In vitro translation of ts NY68 virion RNA at 30.degree. C resulted in efficient synthesis of immunoprecipitable p60src, but very inefficient synthesis of an immunoprecipitable protein kinase. The p60src obtained by in vitro translation of wild-type virion RNA was more than 20-fold more active as a protein kinase than was that obtained from ts Ny68 RNA. The correlation in the case of ts Ny68 of a deficiency in protein kinase activity with an inability to transform cells at high temperature suggests that the protein kinase activity associated with p60src is indeed critical to cellular transformation.