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利用回复细胞系鉴定v-fos转化特异性基因表达改变的靶点。

Use of revertant cell lines to identify targets of v-fos transformation-specific alterations in gene expression.

作者信息

Hoemann C D, Zarbl H

机构信息

Division of Toxicology, Whitaker College, Massachusetts Institute of Technology, Cambridge 02139.

出版信息

Cell Growth Differ. 1990 Dec;1(12):581-90.

PMID:2126949
Abstract

Two proteins expressed in Rat-1 cells which are targets for v-fos transformation-specific alterations in gene expression were identified as alpha 1(I) and alpha 2(I) procollagen. While procollagen (I) proteins were synthesized in Rat-1 fibroblasts, their synthesis was dramatically reduced in Rat-1 cells transformed with the FBJ-v-fos oncogene. Revertant cell lines, which were previously shown to express a functional fos oncoprotein, resumed the synthesis of procollagen (I) at levels comparable to those seen in Rat-1 cells. Further results indicated that these procollagen proteins were also synthesized in Rat-1 cell lines that constitutively express high levels of a transfected c-fos protooncogene. Together, these observations suggested that constitutive fos protein expression was not sufficient to inhibit synthesis of these proteins. We have further demonstrated that Rat-1 cells transformed by most other oncogenes express abundant levels of procollagen (I), indicating that inhibition of procollagen (I) synthesis is not a general characteristic of transformed Rat-1 cells but is specifically associated with FBJ-v-fos-induced transformation. Northern blot analysis and runoff transcription assay data indicated that the alpha 1(I) procollagen, but not alpha 2(I) procollagen, is differentially regulated at the transcriptional level in Rat-1 fibroblasts, v-fos transformants, and revertants.

摘要

在大鼠-1细胞中表达的两种蛋白质被鉴定为α1(I)和α2(I)前胶原,它们是v-fos转化特异性基因表达改变的靶点。虽然前胶原(I)蛋白在大鼠-1成纤维细胞中合成,但在用FBJ-v-fos癌基因转化的大鼠-1细胞中其合成显著减少。先前显示表达功能性fos癌蛋白的回复细胞系恢复了前胶原(I)的合成,其水平与在大鼠-1细胞中所见相当。进一步的结果表明,这些前胶原蛋白也在组成性表达高水平转染c-fos原癌基因的大鼠-1细胞系中合成。总之,这些观察结果表明,组成性fos蛋白表达不足以抑制这些蛋白质的合成。我们进一步证明,被大多数其他癌基因转化的大鼠-1细胞表达大量的前胶原(I),这表明前胶原(I)合成的抑制不是转化的大鼠-1细胞的普遍特征,而是与FBJ-v-fos诱导的转化特异性相关。Northern印迹分析和径流转录测定数据表明,α1(I)前胶原而非α2(I)前胶原在大鼠-1成纤维细胞、v-fos转化体和回复体中在转录水平上受到差异调节。

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