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Abstract
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.
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1 Mahidol University, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); Mahidol University, Department of Biochemistry, Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
2 National Science and Technology Development Agency (NSTDA), National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408)
3 Mahidol University, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); National Science and Technology Development Agency (NSTDA), National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408)
4 Mahidol University, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); National Science and Technology Development Agency (NSTDA), Thailand Science Park, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408)
5 Mahidol University, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); Mahidol University, Department of Anatomy, Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
6 Mahidol University, Department of Biochemistry, Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); Mahidol University, Laboratory of Molecular Cell Biology, Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
7 Chulalongkorn University, Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Bangkok, Thailand (GRID:grid.7922.e) (ISNI:0000 0001 0244 7875)