Guanine nucleotide-binding activity as an assay for src protein of rat-derived murine sarcoma viruses.
Guanine nucleotide-binding activity as an assay for src protein of rat-derived murine sarcoma viruses.
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鸟嘌呤核苷酸结合活性作为大鼠源性鼠肉瘤病毒 src 蛋白的测定。
DOI：
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发表时间：
1979
影响因子：
11.1
通讯作者：
T. Y. Shih
中科院分区：
文献类型：
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作者：
E. Scolnick;A. Papageorge;T. Y. Shih
我们最近发现了一种 21,000 道尔顿的蛋白质 p21，由 Kirsten 或 Harvey 鼠肉瘤病毒编码。根据用对维持转化温度敏感的克尔斯滕肉瘤病毒突变体p21获得的结果，我们得出结论，p21是维持由任一病毒诱导的转化所必需的。我们在此报道，当用[(3)H]GDP或[α-(32)P]GTP孵育Kirsten或Harvey肉瘤病毒转化的细胞的提取物时，皮摩尔量的鸟嘌呤核苷酸可以用含有p21抗体的抗血清进行免疫沉淀。之前我们已经证明克尔斯滕肉瘤病毒温度敏感突变体的[(35)S]甲硫氨酸标记的p21的免疫沉淀性是不耐热的。本文显示鸟嘌呤核苷酸的结合在相同突变体转化的细胞提取物中也是热不稳定的。然而，如果在加热之前将含有标记 p21 的提取物与添加的 GDP 或 GTP 一起温育，则可以保留温度敏感突变体的此类提取物中 [(35)S] 甲硫氨酸标记的 p21 的免疫沉淀性。结果表明p21和某些鸟嘌呤核苷酸之间存在相互作用，并讨论了鸟嘌呤核苷酸和p21在维持转化中的可能作用。
We have recently identified a 21,000-dalton protein, p21, coded for by Kirsten or Harvey murine sarcoma virus. On the basis of the results obtained with the p21 of a mutant of Kirsten sarcoma virus, temperature sensitive for the maintenance of transformation, we concluded that the p21 was required for the maintenance of transformation induced by either virus. We report herein that when extracts from cells transformed by Kirsten or Harvey sarcoma virus are incubated with [(3)H]GDP or [alpha-(32)P]GTP, picomole quantities of guanine nucleotide can be immunoprecipitated with antisera that contain antibodies to the p21. Previously we have shown that the immunoprecipitability of [(35)S]methionine-labeled p21 of the temperature-sensitive mutant of Kirsten sarcoma virus is thermolabile. The binding of guanine nucleotide is shown herein also to be thermolabile in extracts of cells transformed by the same mutant. However, the immunoprecipitability of the [(35)S]methionine-labeled p21 in such extracts of the temperature-sensitive mutant can be preserved if the extracts containing labeled p21 are incubated with added GDP or GTP prior to heating. The results suggest an interaction between p21 and certain guanine nucleotides, and the possible roles of guanine nucleotides and p21 in the maintenance of transformation are discussed.