OVEREXPRESSED PP60C-SRC CAN INDUCE FOCUS FORMATION WITHOUT COMPLETE TRANSFORMATION OF NIH 3T3 CELLS
OVEREXPRESSED PP60C-SRC CAN INDUCE FOCUS FORMATION WITHOUT COMPLETE TRANSFORMATION OF NIH 3T3 CELLS
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DOI：
10.1128/mcb.5.5.1073
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发表时间：
1985-01-01
影响因子：
5.3
通讯作者：
SHALLOWAY, D
中科院分区：
文献类型：
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作者：
JOHNSON, PJ;COUSSENS, PM;SHALLOWAY, D
将含有Moloney小鼠白血病病毒长末端重复序列和鸡c-src或v-src基因的重组质粒分别导入NIH 3T3细胞。与含有c-src和禽类逆转录病毒或SV40启动子增强子的表达载体相比，两种Moloney小鼠白血病病毒长末端重复序列src表达载体均可诱导病灶形成，尽管c-src诱导的病灶数仅为v-src的1%。焦点选择的c-src过表达细胞改变了形态，限制了在软琼脂中的生长，但在体内不会致瘤。克利夫兰消化、体外比较激酶分析、二次转染法和免疫沉淀法表明，病灶的形成是由罕见的导致pp60c-src高水平表达的转基因事件引起的，而不是由转基因c-src基因的突变引起的。Pp60v-src诱导的转化不是完全虚假的活动，与pp60c-src的功能无关，而是代表了涉及pp60c-src的已有分子调控过程的扰动。
NIH 3T3 cells were transfected with plasmids containing Moloney murine leukemia virus long terminal repeats and either chicken c-src or v-src genes. In contrast with the effects observed after transfection with plasmids containing c-src and avian retrovirus or SV40 promoter-enhancers both types of Moloney murine leukemia virus long terminal repeat-src expression plasmids induced focus formation, although c-src induced only 1% as many foci as v-src. The focus-selected c-src overexpressed cells had altered morphology and limited growth in soft agarose but were not tumorigenic in vivo. Cleveland digests, comparative in vitro kinase assays, secondary transfections, and immunoprecipitations indicated that focus formation was caused by rare transfection events that resulted in very high-level pp60c-src expression rather than by mutations of the transfected c-src genes. The pp60v-src induced transformation is not a completely spurious activity which is unrelated to the function of pp60c-src but that it represents a perturbation of already existent molecular control processes involving pp60c-src.