Shaffer J B, Sutton R B, Bewley G C
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201.
J Biol Chem. 1987 Sep 25;262(27):12908-11.
We have investigated the genetic control of murine catalase expression by analyzing catalase transcription and translation products from the tissues of control (Csa) and acatalasemic (Csb) mouse strains. Csb animals possess nearly normal catalase enzyme activity levels in liver, while displaying approximately 20 and 1% of normal activity levels in kidney and red blood cells, respectively. Immunoblot analyses of catalase in these tissues have revealed reduced levels of immunologically reactive catalase protein in Csb kidney and red blood cells, paralleling the reduction of catalase enzyme activity in these tissues. In order to determine the molecular basis for Csb acatalasemia, we have isolated a cDNA clone for murine catalase and have used this probe to analyze Csa and Csb genomic DNA and catalase mRNA. These studies have revealed: 1) no restriction fragment length polymorphisms between Csa and Csb genomic DNAs; 2) no differences in the levels of Csa and Csb catalase mRNA within a single tissue; and 3) no differences in the sizes of Csa and Csb catalase mRNAs. These observations suggest that the genetic defect that produces the tissue-specific reduction of catalase expression in Csb mice is not due to a marked rearrangement of DNA within the Csb catalase structural gene. Furthermore, the Csb mutation does not act at the level of gene transcription or mRNA stability, but rather at the level of mRNA translation and/or catalase protein turnover.
我们通过分析对照(Csa)和无过氧化氢酶血症(Csb)小鼠品系组织中的过氧化氢酶转录和翻译产物,研究了小鼠过氧化氢酶表达的遗传控制。Csb动物肝脏中的过氧化氢酶活性水平几乎正常,而肾脏和红细胞中的活性水平分别约为正常水平的20%和1%。对这些组织中的过氧化氢酶进行免疫印迹分析发现,Csb肾脏和红细胞中具有免疫反应性的过氧化氢酶蛋白水平降低,这与这些组织中过氧化氢酶活性的降低情况相符。为了确定Csb无过氧化氢酶血症的分子基础,我们分离出了小鼠过氧化氢酶的cDNA克隆,并使用该探针分析Csa和Csb的基因组DNA以及过氧化氢酶mRNA。这些研究揭示了:1)Csa和Csb基因组DNA之间不存在限制性片段长度多态性;2)单个组织内Csa和Csb过氧化氢酶mRNA水平没有差异;3)Csa和Csb过氧化氢酶mRNA的大小没有差异。这些观察结果表明,导致Csb小鼠组织特异性过氧化氢酶表达降低的遗传缺陷并非由于Csb过氧化氢酶结构基因内DNA的明显重排。此外,Csb突变并非作用于基因转录或mRNA稳定性水平,而是作用于mRNA翻译和/或过氧化氢酶蛋白周转水平。