Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein
Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein
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DOI：
10.1128/mcb.6.12.4155-4160.1986
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发表时间：
1986-12
影响因子：
5.3
通讯作者：
J. Kato;T. Takeya;C. Grandori;H. Iba;J. Levy;H. Hanafusa
中科院分区：
文献类型：
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作者：
J. Kato;T. Takeya;C. Grandori;H. Iba;J. Levy;H. Hanafusa
我们之前已经证明，携带病毒 src 基因 (v-src) 的细胞同源物 (c-src) 的劳斯肉瘤病毒变种不会转化鸡胚成纤维细胞。我们还表明，用 v-src 的相应区域替换细胞 src 基因 BglI 位点上游或下游的序列可恢复杂合基因的转化活性。由于BglI上游序列中p60c-src和p60v-src之间只有6个氨基酸变化，因此我们构建了涉及v-src和c-src的嵌合分子，以确定每个氨基酸取代对基因产物的生物活性的影响。我们发现，在第 338 位从 Thr 变为 Ile，或者用包含 Asp-63、Trp-95 和 Ile-96 的 v-src 相应片段替换包含 Gly-63、Arg-95 和 Thr-96 的 c-src 片段，根据病灶形成、锚定独立生长和肿瘤形成的标准，将 p60c-src 转化为转化蛋白。 刚出生的小鸡。这些突变还导致 p60c-src 的蛋白激酶活性升高。
We have previously shown that Rous sarcoma virus variants that carry the cellular homolog (c-src) of the viral src gene (v-src) do not transform chicken embryo fibroblasts. We also have shown that replacement of sequences upstream or downstream from the BglI site of the cellular src gene with the corresponding regions of v-src restored transforming activity to the hybrid genes. Since there are only six amino acid changes between p60c-src and p60v-src within the sequences upstream from BglI, we constructed chimeric molecules involving v-src and c-src to determine the effect of each amino acid substitution on the biological activities of the gene product. We found that the change from Thr to Ile at position 338 or the replacement of a fragment of c-src containing Gly-63, Arg-95, and Thr-96 with a corresponding fragment of v-src containing Asp-63, Trp-95, and Ile-96 converted p60c-src into a transforming protein by the criteria of focus formation, anchorage-independent growth, and tumor formation in newborn chickens. These mutations also resulted in elevation of the protein kinase activity of p60c-src.