Norton P A, Hynes R O
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Mol Cell Biol. 1987 Dec;7(12):4297-307. doi: 10.1128/mcb.7.12.4297-4307.1987.
To study the alternative splicing of fibronectin during embryogenesis and oncogenic transformation, we isolated cDNA clones of chicken fibronectin. The partial amino acid sequence deduced from sequencing of these clones showed that, overall, chicken fibronectin is approximately 80% identical with mammalian fibronectins. However, two of the three known regions of alternative splicing differed from this average. The V region was significantly more divergent, and RNA from embryonic chicken liver showed a pattern of V exon splicing which was distinct from that seen in human or rat fibronectins. In contrast, the EIIIB segment was very highly conserved (96%). As in mammals, this segment and another (EIIIA) were alternatively spliced in a cell-type-specific fashion. EIIIA+ and EIIIB+ species were almost absent in liver but predominated in total embryo RNA at all times from 2.5 to 11 days postfertilization. We also examined the possible contributions of fibronectin splicing and integrin receptor expression to the loss of fibronectin on oncogenic transformation. We detected little change in fibronectin splicing, other than a slight increase in representation of EIIIB+ species in fibroblasts after transformation by Rous sarcoma virus. It was also established that the overall reduction in fibronectin mRNA level observed after transformation was not accompanied by a decrease in integrin mRNA levels, indicating that fibronectin and integrin receptors are not coordinately regulated at the transcriptional level.
为了研究胚胎发育和致癌转化过程中纤连蛋白的可变剪接,我们分离了鸡纤连蛋白的cDNA克隆。从这些克隆的测序结果推导的部分氨基酸序列表明,总体而言,鸡纤连蛋白与哺乳动物纤连蛋白的相似度约为80%。然而,三个已知的可变剪接区域中有两个与这个平均值不同。V区域的差异明显更大,来自胚胎鸡肝脏的RNA显示出的V外显子剪接模式与人类或大鼠纤连蛋白中的不同。相比之下,EIIIB片段高度保守(96%)。与哺乳动物一样,该片段和另一个片段(EIIIA)以细胞类型特异性方式进行可变剪接。EIIIA+和EIIIB+类型在肝脏中几乎不存在,但在受精后2.5至11天的整个胚胎RNA中始终占主导地位。我们还研究了纤连蛋白剪接和整合素受体表达对致癌转化过程中纤连蛋白缺失的可能作用。我们检测到纤连蛋白剪接变化很小,只是在劳氏肉瘤病毒转化后的成纤维细胞中EIIIB+类型的比例略有增加。还确定了转化后观察到的纤连蛋白mRNA水平的总体降低并没有伴随着整合素mRNA水平的降低,这表明纤连蛋白和整合素受体在转录水平上没有协同调节。