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果蝇的norpA基因编码多种磷脂酶C亚型。

Multiple subtypes of phospholipase C are encoded by the norpA gene of Drosophila melanogaster.

作者信息

Kim S, McKay R R, Miller K, Shortridge R D

机构信息

Department of Biological Sciences, State University of New York, Buffalo, New York 14260, USA.

出版信息

J Biol Chem. 1995 Jun 16;270(24):14376-82. doi: 10.1074/jbc.270.24.14376.

Abstract

The norpA gene of Drosophila melanogaster encodes a phosphatidylinositol-specific phospholipase C that is essential for phototransduction. Besides being found abundantly in retina, norpA gene products are expressed in a variety of tissues that do not contain phototransduction machinery, implying that norpA is involved in signaling pathways in addition to phototransduction. We have identified a second subtype of norpA protein that is generated by alternative splicing of norpA RNA. The alternative splicing occurs at a single exon that is excluded from mature norpA transcripts when a substitute exon of equal size is retained. The net difference between the two subtypes of norpA protein is 14 amino acid substitutions occurring between amino acid positions 130 and 155 of the enzyme. Results from Northern analyses suggest that norpA subtype I transcripts are most abundantly expressed in adult retina, while subtype II transcripts are most abundant in adult body. Moreover, norpA subtype I RNA can be detected by the reverse transcription-polymerase chain reaction in extracts of adult head tissue but not adult body nor at earlier stages of Drosophila development. Conversely, norpA subtype II RNA can be detected by reverse transcription-polymerase chain reaction throughout development as well as in heads and bodies of adults. Furthermore, norpA subtype I RNA is easily detected in retina using tissue in situ hybridization analysis, while subtype II RNA is not detectable in retina but is found in brain. Since only norpA subtype I RNA is found in retina, we conclude that subtype I protein is utilized in phototransduction. Since norpA subtype II RNA is not found in retina but is expressed in a variety of tissues not known to contain phototransduction machinery, subtype II protein is likely to be utilized in signaling pathways other than phototransduction. The amino acid differences between the two subtypes of norpA protein may reflect the need for each subtype to interact with signaling components of different signal-generating pathways.

摘要

黑腹果蝇的norpA基因编码一种磷脂酰肌醇特异性磷脂酶C,它对光转导至关重要。除了在视网膜中大量存在外,norpA基因产物还在多种不包含光转导机制的组织中表达,这意味着norpA除了参与光转导外,还涉及信号通路。我们已经鉴定出由norpA RNA的可变剪接产生的norpA蛋白的第二种亚型。可变剪接发生在一个单一外显子上,当保留一个等大小的替代外显子时,该外显子被排除在成熟的norpA转录本之外。norpA蛋白的两种亚型之间的净差异是在该酶的第130至155个氨基酸位置之间发生的14个氨基酸替换。Northern分析结果表明,norpA亚型I转录本在成年视网膜中表达最为丰富,而亚型II转录本在成年果蝇身体中最为丰富。此外,通过逆转录-聚合酶链反应可以在成年头部组织提取物中检测到norpA亚型I RNA,但在成年果蝇身体或果蝇发育的早期阶段则检测不到。相反,通过逆转录-聚合酶链反应在整个发育过程以及成年果蝇的头部和身体中都可以检测到norpA亚型II RNA。此外,使用组织原位杂交分析在视网膜中很容易检测到norpA亚型I RNA,而在视网膜中检测不到亚型II RNA,但在脑中可以发现。由于在视网膜中只发现了norpA亚型I RNA,我们得出结论,亚型I蛋白用于光转导。由于在视网膜中未发现norpA亚型II RNA,但在多种已知不包含光转导机制的组织中表达,所以亚型II蛋白可能用于光转导以外的信号通路。norpA蛋白的两种亚型之间的氨基酸差异可能反映了每种亚型与不同信号产生途径的信号成分相互作用的需求。

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