Sanchez M, Gionti E, Arcella A, Pontarelli G, De Lorenzo F
Dipartimento di Biochimica e Biotecnologie Mediche, University of Naples, Italy.
Biochem J. 1993 Oct 1;295 ( Pt 1)(Pt 1):115-9. doi: 10.1042/bj2950115.
alpha 2(I) collagen gene expression is induced in quail embryo chondrocytes pretreated with retinoic acid (RA). The initial appearance of alpha 2(I) mRNA occurs around day 3 of culture in RA-free medium and rapidly progresses over the next 4 days. In transient transfection assays, expression of COL1A2-CAT, a chimeric gene bearing 3500 bp upstream the bone/tendon transcription start site from the human alpha 2(I) gene fused to the CAT gene, is stimulated severalfold in RA-treated chondrocytes. In contrast, enzyme activity is very low in untreated chondrocytes, suggesting that the sequences required for RA-induced transcription of the alpha 2(I) gene are present in this plasmid. Analysis of alpha 2(I) promoter sequences performed with deletion mutants gives overlapping results in collagen type I-producing fibroblasts and chondrocytes withdrawn from RA treatment. These experiments suggest that RA-induced transcription of the alpha 2(I) collagen gene in chondrocytes is regulated by the binding of transcription factors to the same regulatory sequences that control transcription in fibroblasts.
视黄酸(RA)预处理的鹌鹑胚胎软骨细胞中诱导了α2(I)胶原蛋白基因表达。α2(I)mRNA最初出现在无RA培养基培养的第3天左右,并在接下来的4天迅速增加。在瞬时转染试验中,COL1A2-CAT(一种嵌合基因,其携带来自人α2(I)基因的骨/肌腱转录起始位点上游3500 bp与CAT基因融合)在RA处理的软骨细胞中的表达被刺激了数倍。相比之下,未处理的软骨细胞中的酶活性非常低,这表明该质粒中存在RA诱导α2(I)基因转录所需的序列。用缺失突变体对α2(I)启动子序列进行分析,在停止RA处理的I型胶原蛋白产生的成纤维细胞和软骨细胞中得到了重叠的结果。这些实验表明,软骨细胞中RA诱导的α2(I)胶原蛋白基因转录是由转录因子与控制成纤维细胞转录的相同调控序列结合来调节的。