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首页> 外文期刊>Marine genomics >Evolution and comparative genomics of subcellular specializations: EST sequencing of Torpedo electric organ
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Evolution and comparative genomics of subcellular specializations: EST sequencing of Torpedo electric organ

机译：亚细胞专长的演变和比较基因组学：鱼雷电器官的EST测序

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Uncharacterized open reading frames (ORFs) in human genomic sequence often show a high degree of evolutionary conservation, yet have little or no tissue EST or protein data suggestive of protein product function. The encoded proteins may have highly restricted expression in specialized cells, subcellular specializations, and/or narrow windows during development. One such highly specialized and minute subcellular compartment is the neuromuscular junction (NMJ), where motorneurons contact muscle fibers. The electric Torpedo ray has evolved to expand the NMJ structure to the size of a large organ (electroplax organ), and we hypothesized that Torpedo electroplax proteins would be candidates for human ESTs expressed at the human NMJ. A total of 9719 primary electroplax cDNA clones were sequenced. We identified 44 human ORFs showing high (>63%) amino acid identity to Torpedo electroplax transcripts with enrichment for mRNA splicing motifs (SH2 and pre-mRNA splicing domains), an observation potentially important for the strict nuclear domains maintained by myonuclei underlying the NMJ. We generated antibodies against two uncharacterized human genes (C19orf29 [Drosophila cactin] and C15orf24) and showed that these were indeed expressed at the murine NMJ. Cactin, a member of the Rel transcription factor family in Drosophila, localized to the postsynaptic cytosol of the NMJ and nuclear membrane. C15orf24 protein localized to the murine postsynaptic sarcolemma. We show a novel approach towards identifying proteins expressed at a subcellular specialization using evolutionary diversity of organ function and cross-species mapping.
机译：人类基因组序列中未表征的开放阅读框（ORF）通常显示出高度的进化保守性，但几乎没有或没有组织EST或表明蛋白质产物功能的蛋白质数据。在发育过程中，编码的蛋白质在特化细胞，亚细胞特化和/或狭窄窗口中的表达可能受到高度限制。这种高度专业化的微小亚细胞区室之一是神经肌肉接头（NMJ），其中运动神经元接触肌肉纤维。电鱼雷射线已经进化为将NMJ结构扩展到大器官（电浆器官）的大小，并且我们假设鱼雷电浆蛋白将是在人NMJ上表达的人EST的候选物。总共对9719个初级Electroplax cDNA克隆进行了测序。我们鉴定出44个人类ORF，它们与Torpedo电pla转录物表现出高（> 63％）氨基酸同一性，并富集mRNA剪接基序（SH2和pre-mRNA剪接域），这一发现对于NMJ背后的肌核所维持的严格核域可能具有重要意义。我们产生了针对两个未表征的人类基因（C19orf29 [Drosophila cactin]和C15orf24）的抗体，并显示它们确实在鼠NMJ中表达。肌动蛋白，果蝇的Rel转录因子家族的成员，位于NMJ和核膜的突触后细胞质中。 C15orf24蛋白定位于鼠突触后肉瘤。我们展示了一种新的方法来鉴定使用器官功能和跨物种作图进化多样性在亚细胞专业化表达的蛋白质。

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