König H
Kernforschungszentrum Karlsruhe, Institut für Genetik und Toxikologie, FRG.
Nucleic Acids Res. 1991 Jul 11;19(13):3607-11. doi: 10.1093/nar/19.13.3607.
At the serum response element (SRE) of the c-fos proto-oncogene a ternary complex with two proteins, ternary complex factor (p62TCF) and serum response factor (SRF) can be formed. Its formation has previously been suggested to be necessary for efficient induction of c-fos transcription by serum in mouse NIH 3T3 fibroblasts (1) and by phorbol esters, but not by serum, in mouse BALB/c 3T3 fibroblasts (2). It is shown here by genomic dimethyl sulfate (DMS) footprinting that this ternary complex is indeed formed in NIH 3T3 cells in vivo. However, cells were found, murine F9 teratocarcinoma stem cells, in which the pattern of protection and hyperreactivity is consistent with the absence of p62TCF in the protein complex in vivo, although inducibility of the endogenous c-fos gene is not impaired. Both in NIH 3T3 cells and in F9 cells a mutated c-fos promoter that binds SRF but fails to form a ternary complex, was inducible by serum and phorbol esters to the same extent as the wild-type promoter. The data suggest that ternary complex formation is not an absolute prerequisite for the transient induction of c-fos. Ternary complex formation rather appears to enhance overall promoter efficiency. A cell type specific component determines the formation of the multicomponent transcription factor complex in vivo.
在原癌基因c-fos的血清反应元件(SRE)处,可形成一种由两种蛋白质组成的三元复合物,即三元复合物因子(p62TCF)和血清反应因子(SRF)。此前有人提出,在小鼠NIH 3T3成纤维细胞中(1),其形成对于血清有效诱导c-fos转录是必需的;而在小鼠BALB/c 3T3成纤维细胞中(2),对于佛波酯诱导c-fos转录是必需的,但对于血清诱导则不是必需的。本文通过基因组硫酸二甲酯(DMS)足迹法表明,这种三元复合物确实在NIH 3T3细胞体内形成。然而,发现小鼠F9畸胎瘤干细胞体内的蛋白质复合物中存在一种保护和高反应性模式,这与p62TCF的缺失一致,尽管内源性c-fos基因的诱导性并未受损。在NIH 3T3细胞和F9细胞中,一个与SRF结合但无法形成三元复合物的突变型c-fos启动子,被血清和佛波酯诱导的程度与野生型启动子相同。这些数据表明,三元复合物的形成并非c-fos瞬时诱导的绝对先决条件。相反,三元复合物的形成似乎提高了整体启动子效率。一种细胞类型特异性成分决定了体内多组分转录因子复合物的形成。