Labelle Y, Phaneuf D, Tanguay R M
Ontogénèse et Génétique Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université Laval, Sainte-Foy, Québec, Canada.
Gene. 1991 Aug 15;104(2):197-202. doi: 10.1016/0378-1119(91)90250-f.
Fumarylacetoacetate hydrolase (FAH) is an enzyme which is deficient in human hereditary tyrosinemia type 1. We have cloned and sequenced a rat liver cDNA encoding FAH. The identity of the clone was ascertained by hybrid-selection experiments and deduced amino acid (aa) sequence homologies with sequenced oligopeptide fragments of the purified rat liver protein. The cDNA codes for a 419-aa protein of 45,946 daltons. We used this cDNA as a probe in conjunction with a specific anti-rat FAH antibody to study the expression pattern of the FAH gene in rat liver and kidney. Northern blot analysis indicates that the kidney contains slightly more FAH mRNA that the liver. Western blotting shows, however, that the liver contains about twice as much FAH protein as the kidney. Primer extension experiments suggest that there are no differences in the 5'-untranslated (UT) ends of the FAH mRNA of both tissues. We conclude that synthesis of the FAH protein is in part regulated at the post-transcriptional level in rats liver and kidney, and that this regulation does not appear to be mediated by the 5'-UT sequence of the FAH mRNA.
富马酰乙酰乙酸水解酶（FAH）是一种在人类1型遗传性酪氨酸血症中缺乏的酶。我们已经克隆并测序了编码FAH的大鼠肝脏cDNA。通过杂交选择实验确定了该克隆的身份，并通过与纯化的大鼠肝脏蛋白的测序寡肽片段推导的氨基酸（aa）序列同源性进行了验证。该cDNA编码一个45946道尔顿的419个氨基酸的蛋白质。我们使用该cDNA作为探针，并结合特异性抗大鼠FAH抗体来研究FAH基因在大鼠肝脏和肾脏中的表达模式。Northern印迹分析表明，肾脏中含有的FAH mRNA略多于肝脏。然而，Western印迹显示，肝脏中含有的FAH蛋白量约为肾脏的两倍。引物延伸实验表明，两种组织的FAH mRNA的5'-非翻译（UT）末端没有差异。我们得出结论，FAH蛋白的合成在大鼠肝脏和肾脏中部分受转录后水平调控，并且这种调控似乎不是由FAH mRNA的5'-UT序列介导的。
