Ann D K, Wu M M, Huang T, Carlson D M, Wu R
Department of Biochemistry and Biophysics, California Primate Research Center, Davis.
J Biol Chem. 1988 Mar 15;263(8):3546-9.
Conducting airway epithelial cells requires vitamin A or its synthetic chemicals (retinoids) for their survival and for the expression of normal mucociliary functions. By using molecular cloning, we have shown that one of the effects of retinol on cultured human tracheobronchial epithelial (HTBE) cells is the enhancement (from 2- to 4-fold) of the mRNA encoding the elongation factor EF-1 alpha. Sequence analysis has shown that clone HT7, which was identified by differential hybridization procedures, contained a cDNA insert which encoded a protein closely resembling (81%) elongation factor EF-1 alpha from brine shrimp and completely identical to the published sequence of human elongation factor EF-1 alpha (Brands, H.H.G.M., Maassen, J.A., Van Hemert, F.J., Amons, R., and Moller, W. (1986) Eur. J. Biochem. 155, 167-171). Regions of homology of HT7 to EF-Tu from yeast mitochondria, plant chloroplasts, and Escherichia coli are also evident. A single RNA band at 1700 bases was observed for both untreated and retinol-treated HTBE cells, and for mouse liver and parotid glands when Northern transfer from denaturing agarose gel was probed with a 32P-labeled HT7 insert. An enhanced amino acid incorporation and increased protein content per cell for HTBE cells grown in the presence of retinol were observed. Results presented by these studies indicate that retinol may regulate the transcription of a factor required for translation.
气道上皮细胞需要维生素A或其合成化学物质(类视黄醇)来维持生存并表达正常的黏液纤毛功能。通过分子克隆,我们发现视黄醇对培养的人气管支气管上皮(HTBE)细胞的作用之一是增强(2至4倍)编码延伸因子EF-1α的mRNA。序列分析表明,通过差异杂交程序鉴定的克隆HT7包含一个cDNA插入片段,该片段编码的蛋白质与卤虫的延伸因子EF-1α非常相似(81%),并且与已发表的人延伸因子EF-1α序列完全相同(Brands, H.H.G.M., Maassen, J.A., Van Hemert, F.J., Amons, R., and Moller, W. (1986) Eur. J. Biochem. 155, 167 - 171)。HT7与酵母线粒体、植物叶绿体和大肠杆菌的EF-Tu的同源区域也很明显。当用32P标记的HT7插入片段对变性琼脂糖凝胶进行Northern转移杂交时,未处理和视黄醇处理的HTBE细胞以及小鼠肝脏和腮腺均观察到一条1700个碱基的单一RNA条带。在视黄醇存在下生长的HTBE细胞中观察到氨基酸掺入增强且每个细胞的蛋白质含量增加。这些研究结果表明视黄醇可能调节翻译所需因子的转录。