Ohnishi Yoshinori N, Sakumi Kunihiko, Yamazaki Katsuhisa, Ohnishi Yoko H, Miura Tomofumi, Tominaga Yohei, Nakabeppu Yusaku
Division of Neurofunctional Genomics, Department of Immunobiology and Neuroscience, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8582, Japan.
Mol Biol Cell. 2008 Nov;19(11):4717-29. doi: 10.1091/mbc.e07-08-0768. Epub 2008 Aug 27.
Among fos family genes encoding components of activator protein-1 complex, only the fosB gene produces two forms of mature transcripts, namely fosB and DeltafosB mRNAs, by alternative splicing of an exonic intron. The former encodes full-length FosB. The latter encodes DeltaFosB and Delta2DeltaFosB by alternative translation initiation, and both of these lack the C-terminal transactivation domain of FosB. We established two mutant mouse embryonic stem (ES) cell lines carrying homozygous fosB-null alleles and fosB(d) alleles, the latter exclusively encoding DeltaFosB/Delta2DeltaFosB. Comparison of their gene expression profiles with that of the wild type revealed that more than 200 genes were up-regulated, whereas 19 genes were down-regulated in a DeltaFosB/Delta2DeltaFosB-dependent manner. We furthermore found that mRNAs for basement membrane proteins were significantly up-regulated in fosB(d/d) but not fosB-null mutant cells, whereas genes involved in the TGF-beta1 signaling pathway were up-regulated in both mutants. Cell-matrix adhesion was remarkably augmented in fosB(d/d) ES cells and to some extent in fosB-null cells. By analyzing ES cell lines carrying homozygous fosB(FN) alleles, which exclusively encode FosB, we confirmed that FosB negatively regulates cell-matrix adhesion and the TGF-beta1 signaling pathway. We thus concluded that FosB and DeltaFosB/Delta2DeltaFosB use this pathway to antagonistically regulate cell matrix adhesion.
在编码激活蛋白-1复合物组分的fos家族基因中,只有fosB基因通过外显子内含子的可变剪接产生两种成熟转录本,即fosB和DeltafosB mRNA。前者编码全长FosB。后者通过可变翻译起始编码DeltaFosB和Delta2DeltaFosB,并且这两者都缺乏FosB的C末端反式激活结构域。我们建立了两个携带纯合fosB无效等位基因和fosB(d)等位基因的突变小鼠胚胎干细胞(ES)系,后者专门编码DeltaFosB/Delta2DeltaFosB。将它们的基因表达谱与野生型进行比较,发现超过200个基因上调,而19个基因以DeltaFosB/Delta2DeltaFosB依赖的方式下调。我们还发现,在fosB(d/d)突变细胞中基底膜蛋白的mRNA显著上调,但在fosB无效突变细胞中没有,而参与TGF-β1信号通路的基因在两种突变体中均上调。fosB(d/d) ES细胞中的细胞-基质黏附显著增强,在fosB无效细胞中也有一定程度的增强。通过分析携带纯合fosB(FN)等位基因(专门编码FosB)的ES细胞系,我们证实FosB负向调节细胞-基质黏附和TGF-β1信号通路。因此,我们得出结论,FosB和DeltaFosB/Delta2DeltaFosB利用该通路拮抗调节细胞基质黏附。