Echinococcus granulosus tropomyosin isoforms: From gene structure to expression analysis
Echinococcus granulosus tropomyosin isoforms: From gene structure to expression analysis
复制标题
DOI：
10.1016/j.gene.2008.11.021
复制
发表时间：
2009-03-15
期刊：
影响因子：
3.5
通讯作者：
Esteves, Adriana
中科院分区：
文献类型：
--
作者：
Alvite, Gabriela;Esteves, Adriana
原肌球蛋白 (Trps) 是所有真核细胞中发现的肌动蛋白丝结合蛋白家族。在肌肉细胞中，它们通过调节肌动蛋白和肌球蛋白的钙敏感性相互作用在收缩中发挥核心作用。在非肌肉细胞中，原肌球蛋白调节肌动蛋白丝的组织和动力学。 Trps 基因表现出由选择性剪接产生的广泛的细胞类型特异性亚型多样性。在这里，我们报告了寄生扁形动物细粒棘球绦虫的原肌球蛋白基因转录序列的特征。使用RT-PCR方法，我们分离了三种亚型（egtrpA、egtrpB和egtrpC），它们与不同系统发育起源的原肌球蛋白表现出显着的同源性。相应的基因egtrp（5656 bp）包含八个内含子和九个外显子。 Southern 印迹杂交研究表明，egtrp 在细粒棘球绦虫中以单拷贝基因座的形式存在。我们证明egtrp表达三种不同的转录本，它们的不同之处在于可变剪接的外显子4和内含子VI。有趣的是，内含子 VI 遭受内含子保留并在框架中包含内部终止密码子。使用特定的抗 rEgTrp 抗血清还通过蛋白质印迹分析检测到三个主要条带。免疫定位和原位杂交研究表明，egtrp 转录和翻译主要定位于原头节吸盘。这是该寄生虫中选择性剪接的第一份报告。 (C) 2008 Elsevier B.V. 保留所有权利。
Tropomyosins (Trps) constitute a family of actin filament-binding proteins found in all eukaryotic cells. In muscle cells, they play a central role in contraction by regulating calcium-sensitive interaction of actin and myosin. In non-muscle cells, tropomyosins regulate actin filament organization and dynamics. Trps genes exhibit extensive cell type-specific isoform diversity generated by alternative splicing. Here, we report the characterization of tropomyrosin gene transcribed sequences from the parasitic platyhelminth Echinococcus granulosus. Using RT-PCR approach we isolated three isoforms (egtrpA, egtrpB and egtrpC), which display significant homologies to know tropomyosins of different phylogenetic origin. The corresponding gene, egtrp (5656 bp), contains eight introns and nine exons. Southern blot hybridization studies showed that egtrp is present as single copy locus in E. granulosus. We demonstrated that egtrp expresses three different transcripts which differ in alternatively spliced exon 4 and intron VI. Interestingly, intron VI suffers intron retention and contains an internal stop codon in frame. Three major bands are also detected by Western blot analysis using a specific anti-rEgTrp antiserum. Immune-localization and in situ hybridization studies showed that egtrp transcription and translation is mostly localized at the protoscoleces suckers. This is the first report of alternative splicing in this parasite. (C) 2008 Elsevier B.V. All rights reserved.