Matsuura T, Hoshide R, Komaki S, Kiwaki K, Endo F, Nakamura S, Jitosho T, Matsuda I
Department of Pediatrics, Kumamoto University School of Medicine, Japan.
J Inherit Metab Dis. 1995;18(3):273-82. doi: 10.1007/BF00710415.
Ornithine carbamoyltransferase (OCT) is a liver-specific enzyme located in the mitochondrial matrix. OCT deficiency is an X-linked disease with a heterogeneous phenotype, even in affected males. We studied two male patients (K.M., K.G.) with early and late onset, respectively. OCT activity was zero in the autopsied liver of patient K.M. and was 6% of control in the biopsied liver of K.G. Sequencing of OCT cDNAs revealed exon 5 skipping in K.M., resulting from a T-to-C transition of the initial dinucleotide of the 5' splicing donor site of intron 5, and a G-to-T transversion at position +45 in exon 9 (L304F) in K.G., providing three OCT mRNAs of different lengths: a normally spliced transcript, 23 bp insertion of intron 8 and the first 50bp missing within exon 9. Exon 5 skipping and two other aberrant splicings produced stop codons early downstream in mature OCT mRNAs. Expression study of a missense allele, L304F, transfected to cultured Cos 1 cells revealed a 34.4% value of the control. The difference of OCT activities between the patient liver and transfected cells (6% vs. 34%) can be explained by this splicing abnormality.
鸟氨酸氨甲酰基转移酶(OCT)是一种位于线粒体基质中的肝脏特异性酶。OCT缺乏症是一种X连锁疾病,即使在受影响的男性中也具有异质性表型。我们分别研究了两名发病早和发病晚的男性患者(K.M.和K.G.)。在患者K.M.的尸检肝脏中OCT活性为零,而在K.G.的活检肝脏中为对照的6%。OCT cDNA测序显示,K.M.中存在外显子5跳跃,这是由内含子5的5'剪接供体位点起始二核苷酸的T到C转换引起的;在K.G.中,外显子9的第45位发生了G到T的颠换(L304F),产生了三种不同长度的OCT mRNA:一种正常剪接的转录本、内含子8的23 bp插入以及外显子9内缺失的前50 bp。外显子5跳跃和其他两种异常剪接在成熟OCT mRNA的下游早期产生了终止密码子。将错义等位基因L304F转染到培养的Cos 1细胞中的表达研究显示其活性为对照的34.4%。患者肝脏和转染细胞之间OCT活性的差异(6%对34%)可以用这种剪接异常来解释。