Identification and characterization of Clorf36, a transcript highly expressed in photoreceptor cells, and mutation analysis in retinitis pigmentosa
Identification and characterization of Clorf36, a transcript highly expressed in photoreceptor cells, and mutation analysis in retinitis pigmentosa
复制标题
DOI：
10.1016/s0006-291x(03)01410-4
复制
发表时间：
2003-08-29
影响因子：
3.1
通讯作者：
Banfi, S
中科院分区：
文献类型：
--
作者：
Lavorgna, G;Lestingi, M;Banfi, S
通过计算方法，我们确定了一个未表征的人类转录物，染色体I开放阅读框36 (Clorf36)，在视网膜中表达，映射到1q32.3。该cDNA包含一个585 bp的开放阅读框，编码195个氨基酸的蛋白，预测质量为22.7 kDa。在视网膜母细胞瘤细胞系中，还发现了一个编码截断肽的选择性剪接转录物。利用几种来源的人cDNA进行的PCR实验表明，Clorf36在视网膜中优先表达。因此，在原位杂交实验中，以小鼠Clorf36 cDNA片段为探针，在小鼠视网膜成体切片上检测到杂交信号。Clorf36蛋白与小家鼠和红河豚的推定蛋白具有同源性，表明其功能具有进化保守性。对Clorf36基因产物的额外序列分析预测了其亚细胞线粒体定位以及进化保守的磷酸化位点和采用线圈状构象的区域的存在。我们还定义了该基因的基因组结构。这使我们能够对大约300名视网膜色素变性患者的Clorf36编码区进行突变分析。本分析未发现病理突变。(C) 2003 Elsevier Inc.版权所有。
By means of computational methods, we identified an uncharacterized human transcript, Chromosome I open reading frame 36 (Clorf36), that is expressed in the retina and that maps to 1q32.3. The cDNA contains an open reading frame of 585 bp that encodes a 195-aminoacid protein with a predicted mass of 22.7 kDa. An alternatively spliced transcript in a retinoblastoma cell line, encoding for a truncated peptide, was also identified. PCR experiments performed using human cDNA from several sources indicate that Clorf36 has a preferential expression in the retina. Accordingly, in situ hybridization experiments, performed using as probe a murine Clorf36 cDNA fragment, detected a hybridization signal on mouse retinal adult sections. The Clorf36 protein shares homology with putative proteins in Mus musculus and Fugu rubripes, suggesting evolutionary conservation of its function. Additional sequence analysis of the Clorf36 gene product predicts its subcellular mitochondrial localization and the presence of both evolutionary conserved phosphorylation sites and regions adopting a coiled-coil conformation. We also defined the genomic structure of the gene. This enabled us to perform a mutational analysis of the Clorf36 coding region of about 300 patients affected by retinitis pigmentosa. No pathological mutations were detected in this analysis. (C) 2003 Elsevier Inc. All rights reserved.