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Molecular and cellular characterization of four putative nucleotide transporters from the shrimp microsporidian Enterocytozoon hepatopenaei (EHP)
Scientific Reports ( IF 3.8 ) Pub Date : 2023-11-16 , DOI: 10.1038/s41598-023-47114-8
Orawan Thepmanee 1, 2 , Natthinee Munkongwongsiri 3 , Anuphap Prachumwat 1, 3 , Vanvimon Saksmerprome 1, 4 , Sarocha Jitrakorn 1, 4 , Kallaya Sritunyalucksana 3 , Rapeepun Vanichviriyakit 1, 5 , Sittinan Chanarat 2, 6 , Pattana Jaroenlak 7 , Ornchuma Itsathitphaisarn 1, 2
Affiliation  

Microsporidia are obligate intracellular parasites that lost several enzymes required in energy production. The expansion of transporter families in these organisms enables them to hijack ATP from hosts. In this study, nucleotide transporters of the microsporidian Enterocytozoon hepatopenaei (EHP), which causes slow growth in economically valuable Penaeus shrimp, were characterized. Analysis of the EHP genome suggested the presence of four putative nucleotide transporter genes, namely EhNTT1, EhNTT2, EhNTT3, and EhNTT4. Sequence alignment revealed four charged amino acids that are conserved in previously characterized nucleotide transporters. Phylogenetic analysis suggested that EhNTT1, 3, and 4 were derived from one horizontal gene transfer event, which was independent from that of EhNTT2. Localization of EhNTT1 and EhNTT2 using immunofluorescence analysis revealed positive signals within the envelope of developing plasmodia and on mature spores. Knockdown of EhNTT2 by double administration of sequence specific double-stranded RNA resulted in a significant reduction in EHP copy numbers, suggesting that EhNTT2 is crucial for EHP replication in shrimp. Taken together, the insight into the roles of NTTs in microsporidian proliferation can provide the biological basis for the development of alternative control strategies for microsporidian infection in shrimp.



中文翻译:

虾微孢子虫肝对虾肠细胞虫 (EHP) 四种假定核苷酸转运蛋白的分子和细胞特征

微孢子虫是专性细胞内寄生虫,丢失了能量生产所需的几种酶。这些生物体中转运蛋白家族的扩展使它们能够劫持宿主的 ATP。在这项研究中,对导致具有经济价值的对虾生长缓慢的微孢子虫肝对虾肠细胞虫(EHP)的核苷酸转运蛋白进行了表征。EHP 基因组分析表明存在四个假定的核苷酸转运蛋白基因,即 EhNTT1、EhNTT2、EhNTT3 和 EhNTT4。序列比对揭示了先前表征的核苷酸转运蛋白中保守的四个带电氨基酸。系统发育分析表明EhNTT1、3和4源自一个水平基因转移事件,该事件独立于EhNTT2。使用免疫荧光分析对 EhNTT1 和 EhNTT2 进行定位,揭示了发育中的疟原虫的包膜内和成熟孢子上的阳性信号。通过两次施用序列特异性双链RNA来敲低EhNTT2导致EHP拷贝数显着减少,这表明EhNTT2对于虾中的EHP复制至关重要。总之,深入了解 NTT 在微孢子虫增殖中的作用可以为开发对虾微孢子虫感染的替代控制策略提供生物学基础。

更新日期:2023-11-17
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