Weller A, Beck S, Ekblom P
Friedrich-Miescher-Laboratorium der Max-Planck-Gesellschaft, Tübingen, Germany.
J Cell Biol. 1991 Jan;112(2):355-62. doi: 10.1083/jcb.112.2.355.
We have isolated cDNA clones for mouse tenascin and analyzed expression of tenascin mRNAs during embryonic development of the kidney and gut. The deduced amino acid sequence of the mouse tenascin cDNAs shows a modular structure of repeats similar to chicken and human tenascin. In mouse there are 14.5 cysteine-rich repeats with similarity to the EGF repeat, followed by several repeats with similarity to the type III repeat of fibronectin. A longer variant contains 13 fibronectin type III repeats, whereas a shorter splice variant of mouse tenascin lacks the 5 type III repeats that occur directly after the fifth repeat in the longer variant. Contrary to the chicken and human sequences, mouse tenascin does not contain an RGD sequence in the third type III repeat implicated in cell attachment, or in any other positions. In Northern hybridizations to RNA from primary embryonic fibroblasts, the cDNA clone M 20/1 detects two mRNAs with sizes close to 6 and 8 kb. This, and the other data presented here suggest that the two major mouse tenascin polypeptides arise through an alternative RNA splicing. The two major mRNAs are differentially expressed during development. The 8-kb mRNA is more prominent than the 6-kb mRNA throughout prenatal kidney development, but during postnatal development the ratio of the two mRNAs changes. A different expression pattern is seen in the developing gut where the 6-kb mRNA predominates during embryogenesis with the 8-kb mRNA appearing later. The mRNA data of the developing gut correspond with previous protein data, which showed that the shorter Mr 210,000 polypeptide predominates during earlier developmental stages and the larger Mr 260,000 polypeptide appears later in the embryonic gut (Aufderheide, E., and P. Ekblom. 1988. J. Cell Biol. 107:2341-2349).
我们已分离出小鼠肌腱蛋白的cDNA克隆,并分析了肌腱蛋白mRNA在肾脏和肠道胚胎发育过程中的表达情况。从小鼠肌腱蛋白cDNA推导的氨基酸序列显示出一种与鸡和人肌腱蛋白相似的重复模块结构。在小鼠中,有14.5个与EGF重复序列相似的富含半胱氨酸的重复序列,接着是几个与纤连蛋白III型重复序列相似的重复序列。一个较长的变体含有13个纤连蛋白III型重复序列,而小鼠肌腱蛋白的一个较短剪接变体则缺少较长变体中紧接在第五个重复序列之后出现的5个III型重复序列。与鸡和人的序列不同,小鼠肌腱蛋白在参与细胞黏附的第三个III型重复序列或任何其他位置都不包含RGD序列。在对原代胚胎成纤维细胞RNA进行的Northern杂交中,cDNA克隆M 20/1检测到两种大小接近6 kb和8 kb的mRNA。本文呈现的这一结果及其他数据表明,两种主要的小鼠肌腱蛋白多肽是通过选择性RNA剪接产生的。这两种主要的mRNA在发育过程中差异表达。在整个产前肾脏发育过程中,8 kb的mRNA比6 kb的mRNA更突出,但在出生后发育过程中,这两种mRNA的比例发生了变化。在发育中的肠道中观察到不同的表达模式,其中6 kb的mRNA在胚胎发生过程中占主导,8 kb的mRNA随后出现。发育中肠道的mRNA数据与先前的蛋白质数据一致,先前的数据表明,较短的210,000 Mr多肽在发育早期阶段占主导,而较大的260,000 Mr多肽在胚胎肠道中较晚出现(奥夫德海德,E.,和P. 埃克布洛姆。1988年。《细胞生物学杂志》107:2341 - 2349)。