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猴气管支气管上皮细胞中hnRNP A1亚型的cDNA克隆及其视黄醇调控作用

cDNA cloning of a hnRNP A1 isoform and its regulation by retinol in monkey tracheobronchial epithelial cells.

作者信息

An G, Wu R

机构信息

California Primate Research Center, University of California-Davis 95616.

出版信息

Biochim Biophys Acta. 1993 Mar 20;1172(3):292-300. doi: 10.1016/0167-4781(93)90216-z.

Abstract

An isoform of the hnRNP A1 was cloned from a cDNA library of monkey tracheobronchial epithelial (TBE) cells by differential hybridization. The cDNA clone MT77 has an insert of 1756 base pairs and the DNA sequence shares high homology to both human A1 alpha-type and beta-type isoforms with the exception of several differences in the coding and noncoding regions. Like the other two isoforms, MT77 has two polyadenylation sites. A probe prepared from the MT77 clone hybridizes to two message bands at 1.4 and 1.8 kb. Both messages were found in a polysomal preparation, suggesting that both messages are used in A1 protein synthesis. The expression of the A1 gene in monkey TBE cells is stimulated by vitamin A (retinol). The results of nuclear run-on transcriptional assays suggest that this stimulation occurs at the transcriptional level. Furthermore, this effect is not prevented, but superinduced, by cycloheximide. These results suggest that vitamin A may be directly involved in regulating A1 transcription through a mechanism similar to the interactions between the retinoic acid responsive elements and the nuclear receptors of retinoic acid.

摘要

通过差异杂交从猴气管支气管上皮(TBE)细胞的cDNA文库中克隆了hnRNP A1的一种同工型。cDNA克隆MT77有一个1756个碱基对的插入片段,其DNA序列与人类A1α型和β型同工型具有高度同源性,只是在编码区和非编码区存在一些差异。与其他两种同工型一样,MT77有两个聚腺苷酸化位点。从MT77克隆制备的探针与1.4 kb和1.8 kb的两条信使带杂交。在多核糖体制剂中发现了这两条信使,表明这两条信使都用于A1蛋白的合成。维生素A(视黄醇)可刺激猴TBE细胞中A1基因的表达。核转录活性分析结果表明,这种刺激发生在转录水平。此外,环己酰亚胺不会阻止这种效应,反而会使其超诱导。这些结果表明,维生素A可能通过一种类似于视黄酸反应元件与视黄酸核受体之间相互作用的机制直接参与调节A1转录。

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