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红海束毛藻中编码多个新型内含肽的分裂dnaE基因。

Split dnaE genes encoding multiple novel inteins in Trichodesmium erythraeum.

作者信息

Liu Xiang-Qin, Yang Jing

机构信息

Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, Nova Scotia B3H 4H7, Canada.

出版信息

J Biol Chem. 2003 Jul 18;278(29):26315-8. doi: 10.1074/jbc.C300202200. Epub 2003 May 24.

Abstract

Three inteins were found when analyzing a pair of split dnaE genes encoding the catalytic subunit of DNA polymerase III in the oceanic N2-fixing cyanobacterium Trichodesmium erythraeum. The three inteins (DnaE-1, DnaE-2, and DnaE-3) were clustered in a 70-amino acid (aa) region of the predicted DnaE protein. The DnaE-1 intein is 1258 aa long and three times as large as a typical intein, due to the presence of large tandem repeats in which a 57-aa sequence is repeated 17 times. The DnaE-2 intein has a more typical size of 428 aa with putative protein splicing and endonuclease domains. The DnaE-3 intein is a split intein consisting of a 102-aa N-terminal part and a 36-aa C-terminal part encoded on the first and second split dnaE genes, respectively. Synthesis of a mature DnaE protein is predicted to involve expression of two split dnaE genes followed by two protein cis-splicing reactions and one protein trans-splicing reaction. Tandem repeats in the DnaE-1 intein inhibited the protein splicing activity of this intein when tested in Escherichia coli cells and may potentially regulate DnaE synthesis in vivo.

摘要

在分析海洋固氮蓝藻红海束毛藻中一对编码DNA聚合酶III催化亚基的分裂dnaE基因时,发现了三种内含肽。这三种内含肽(DnaE-1、DnaE-2和DnaE-3)聚集在预测的DnaE蛋白的一个70个氨基酸(aa)区域内。DnaE-1内含肽长1258个氨基酸,由于存在大的串联重复序列,其中一个57个氨基酸的序列重复了17次,因此其大小是典型内含肽的三倍。DnaE-2内含肽的大小更典型,为428个氨基酸,具有推定的蛋白质剪接和内切核酸酶结构域。DnaE-3内含肽是一种分裂内含肽,分别由位于第一个和第二个分裂dnaE基因上的102个氨基酸的N端部分和36个氨基酸的C端部分组成。预计成熟DnaE蛋白的合成涉及两个分裂dnaE基因的表达,随后是两个蛋白质顺式剪接反应和一个蛋白质反式剪接反应。当在大肠杆菌细胞中进行测试时,DnaE-1内含肽中的串联重复序列抑制了该内含肽的蛋白质剪接活性,并可能在体内潜在地调节DnaE的合成。

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