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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2014, Vol. 40 Issue (1): 9-15    DOI: 10.3785/j.issn.1008-9209.2013.07.182
生物科学与技术     
鸡硒蛋白T的硒代半胱氨酸插入序列元件、蛋白结构与功能及组织表达差异
葛延松， 曹嫦妤， 王丽丽， 李楠， 江秀清， 李金龙*
(东北农业大学动物医学学院，哈尔滨150030)
Analysis of selenocysteine insertion sequence element， structures and functions and expression profiles of selenoprotein T  in chicken
Ge Yansong, Cao Changyu, Wang Lili, Li Nan, Jiang Xiuqing, Li Jinlong*
(College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China)
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摘要： 应用生物软件分析鸡和其他11种脊椎动物硒蛋白T(selenoprotein T,SelT)的硒代半胱氨酸插入序列(selenocysteine insertion sequence, SECIS)元件、SelT核苷酸和氨基酸序列的同源性，并分析鸡SelT的结构及功能；采用实时荧光定量PCR(fluorescent quantitative real-time PCR, fqRT-PCR)方法检测SelT基因在35日龄鸡体内30种组织中的表达谱。结果显示：脊椎动物SelT的SECIS元件均属于Ⅱ型结构；鸡SelT核苷酸序列与其他11种脊椎动物的同源性在48.0%~85.1%之间，而氨基酸序列与非洲爪蟾、斑马鱼的同源性低于90.0%，与其他9种动物的同源性在90.6%~94.9%之间；鸡SelT属于跨膜蛋白，存在信号肽，属于RDx家族，酶活性分类为EC 2.5.1.18，具有氧化还原功能，且存在Ca2+结合位点。SelT在鸡各组织中广泛表达，在睾丸中含量极其丰富，提示鸡SelT在雄性生殖系统中可能发挥功能。
Abstract: Selenium (Se) was an important, biologically necessary trace element, which played an important biological roles in avian growth, reproduction, immune function and disease resistance. The biological significance of Se was attributed to its occurrence in selenoproteins in the form of selenocysteine (Sec) with a 21 amino acid genetic code. Se uses the stop codon UGA as the coding codon encoding Sec. Selenoproteins involved in many life processes such as antioxidant defense, cell signal transduction, metabolic pathway, development, immune function, and hormone regulates, and it was related to the occurrence and mechanism of many diseases. Selenoprotein T (SelT) was originally identified through silico studies, cloned and expressed in a mammalian cell line, confirmed as a selenoprotein. Previous studies have proved that SelT mainly occurred in Golgi apparatus and endoplasmic reticulum, with important biological function. SelT can act on the Ca2+ homeostasis, involved in neuroendocrine secretion. SelT has also been showed in alter cell adhesion. Up to now, 25 selenoprotein-encoding genes were identified in birds, however, only about half have been functionally characterized, most of which were involved in redox reactions. SelT was a Se-containing protein whose cellular function has not been characterized. Research on the structure and function of SelT was still in its infancy, but reports in mammals about SelT and the information about avian SelT were not clear. The purpose of this study is to explore the selenocysteine insertion sequence (SECIS) element, structures and functions of chicken SelT and the expression profiles of SelT in chicken tissues.
The chicken SelT sequence was used in the experiment. The SECIS elements of 12 vertebrates were analyzed by the SECISearch 2.19. The homologies of the molecule nucleotide and amino acid sequences on vertebrates were analyzed by  DNAStar. The structure and functions of chicken SelT were predicted with the bioinformatics. The distribution of SelT in 30 tissues of 35-day-old chicken was analyzed by  fluorescent quantitative real-time PCR (qRT-PCR).
The results showed that SECIS element of vertebrates belonged  to the type Ⅱ SECIS element. Compared with chicken and other 11 vertebrates, the nucleotide sequence homologies of SelT were 48.0%85.1%. The homologies of amino acid sequences of chicken SelT with Xenopus laevis or Danio rerio were less than 90.0% and with other nine kinds of animals were among 90.6%94.9%. The chicken SelT was a transmembrane protein and had a signal peptide. It belonged to the RDx families and its enzyme classification was EC 2.5.1.18. SelT had binding sites with Ca2+. The results of fqRT-PCR showed that the SelT was widely expressed in chicken tissues, and the expression level in testis was the highest.
In summary, the SECIS element of vertebrate SelT belongs to the type Ⅱ and SelT is highly conserved in vertebrates. Avian SelT contains a CxxU motif in a thioredoxin-like fold, with glutathione S-transferase activity. SelT has binding sites with Ca2+, it was predicted that avian SelT could regulate the Ca2+ homeostasis. Avian SelT is widely expressed in chicken tissues, and the testis has the highest expression level, suggesting that the chicken SelT has a special function in the male reproductive organs. This study provides a basis for further experimental analysis of the structure-function of avian SelT.
出版日期: 2014-01-20
CLC:  Q 956  
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葛延松
曹嫦妤
王丽丽
李楠
江秀清
李金龙*

引用本文:

葛延松， 曹嫦妤， 王丽丽， 李楠， 江秀清， 李金龙*. 鸡硒蛋白T的硒代半胱氨酸插入序列元件、蛋白结构与功能及组织表达差异[J]. 浙江大学学报(农业与生命科学版), 2014, 40(1): 9-15.

Ge Yansong, Cao Changyu, Wang Lili, Li Nan, Jiang Xiuqing, Li Jinlong*. Analysis of selenocysteine insertion sequence element， structures and functions and expression profiles of selenoprotein T  in chicken. Journal of Zhejiang University (Agriculture and Life Sciences), 2014, 40(1): 9-15.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2013.07.182    或    http://www.zjujournals.com/agr/CN/Y2014/V40/I1/9

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