Johnson D W, Speier S, Qian W H, Lane S, Cook A, Suzuki K, Daniel P, Boustany R M
Department of Pediatric Neurology, Duke University Medical Center, Durham, North Carolina, USA.
Am J Med Genet. 1995 Jun 5;57(2):350-60. doi: 10.1002/ajmg.1320570250.
The role of subunit-9 of mitochondrial ATP synthase in Batten disease was defined by characterizing the expression of genes encoding this protein in human tissues. Two genetically distinct neuronal ceroid-lipofuscinoses (NCL) comprise Batten disease: the late-infantile (LINCL) and juvenile (JNCL) types. We tested cell lines and tissues from both types of patients, along with normal controls. Differences in expression between diseased and normal samples were found for both mRNA and protein. Antibody staining of subunit-9 protein was detected in LINCL and JNCL tissues, and in 6 LINCL and 4 of 5 JNCL fibroblast lines. No immunoreactivity was seen in fibroblasts from obligate carriers, normal controls, and 6 other storage disease controls, with the exception of faint staining in Niemann-Pick, type C cells. There was an appreciable difference in staining pattern in both tissue sections and fibroblasts between LINCL and JNCL. Three subunit-9 transcripts (Hum1, Hum2, and Hum3) were specifically detected in NCL and normal human tissue from heart, liver, brain, muscle, and pancreas. Transcriptional regulation of subunit-9 genes was found to be altered in Batten disease. Pseudogenes related to each of the subunit-9 genes were isolated. Sequence analysis of cDNAs spanning the protein-coding regions of the Hum1, Hum2, and Hum3 genes showed conclusively that the primary defect(s) causing NCL are not mutations in the protein-coding regions of the 3 known subunit-9 genes.
通过对编码该蛋白的基因在人体组织中的表达进行表征,确定了线粒体ATP合酶亚基9在巴滕病中的作用。巴滕病包括两种基因不同的神经元蜡样脂褐质沉积症(NCL):晚发性婴儿型(LINCL)和青少年型(JNCL)。我们检测了这两种类型患者的细胞系和组织,以及正常对照。在患病样本和正常样本之间,mRNA和蛋白质的表达均存在差异。在LINCL和JNCL组织以及6个LINCL和成纤维细胞系以及5个JNCL中的4个中检测到亚基9蛋白的抗体染色。在纯合子携带者、正常对照和其他6种储存疾病对照的成纤维细胞中未观察到免疫反应性,除了尼曼-皮克C型细胞中有微弱染色。LINCL和JNCL在组织切片和成纤维细胞中的染色模式存在明显差异。在NCL以及来自心脏、肝脏、大脑、肌肉和胰腺的正常人体组织中特异性检测到三种亚基9转录本(Hum1、Hum2和Hum3)。发现巴滕病中亚基9基因的转录调控发生了改变。分离出了与每个亚基9基因相关的假基因。对跨越Hum1、Hum2和Hum3基因蛋白质编码区的cDNA进行序列分析,最终表明导致NCL的主要缺陷不是3个已知亚基9基因蛋白质编码区的突变。