The Functions Of ATP Synthase And Its Inhibitory Factor In Shrimp Virus Immune Reaction

ATP from mitochondrial ATP synthase was the majority energy sources of live cell activities.ATP synthase is essential for the survival of eukaryotes.But till now,the exact roles of ATP synthase in shrimp innate immunity is still unclear.The paper focuses on studying the function of four mitochondrial ATP synthase related smORFs in viral immunity of M.japonicus.ATP synthase inhibitor 1((MjATPIF1)was hijacked by white spot syndrome virus(WSSV)to promote its replicationMjATPIF1 is a natural ATP synthase inhibitor.It could change the activity of ATP synthase to regulate the life activity of cells.However.it is still unclear about the roles of MjATPIF1 in immune reaction.In this study,we identified a ATP synthase inhibit factor 1 in M.japonicus,and named as MjATPIF1.The full open reading frame(ORF)of MjATPIF1 contains 315 nucleotides,encoding 104 amino acids.MjATPIFl was widely distributed in some immune tissues of M.japonicus at the transcriptional and the protein level.The expression pattern analysis showed that MjATPIFl could be upregulated by WSSV challenge in hemocytes and intestine.RNA interference and protein overexpression(recombinant protein injection)assay showed that MjATPIF1 was beneficial for white spot syndrome virus(WSSV)replication in shrimp.Mitochoncdrial reactive oxygen species(ROS)assay suggested that MjATPIF1 could promote the production of superoxide(SOD).MiATPIF1 could increase the NF-κB activity by producing SOD to promote the WSSV replication.ATP assay showed that MjATPIF1 promoted viral replication by changing the type of cellular respiration.All experimental results demonstrated that MjATPIF1 played a crucial role in promoting WSSV replication of shrimp.ATP synthase inhibitor1(MjATPIF1)was hijacked by WSSV to promote its replication.Three ATP synthase subunits were beneficial to WSSV replication in kuruma shrimpThree smORFs were screened and identified from kuruma shrimp M.japonicus,and named as ATP synthase subunit E(MjATP51),ATP synthase subunit G(MjATP5L)and ATP synthase subunit(MjATP5E),and all of them were the subunits of the F1F0-ATP synthase.They constitute the F1F0-ATP synthase together with the other ATP synthase subunits.ATP synthase could synthesize ATP with proton gradient potential in the oxidative phosphorylation pathway.However,it was unclear about the function of three ATP synthase subunits.MjATP51,MjATP5L and MjATP5E,in the defense reaction of shrimp.In this study,we studied the functions of three ATP svnthase subunits MjATP5I,MjATP5L.and MjATP5E in the viral immunity of kuruma shrimp.The full ORF of MjjATP5Icontained 258 nucletides,encoding a protein of 85 amino acids.The sequence of MjATP5L was 300 bp in length,encoding 99 amino acids.The ORF of MjATP5E was 168 bp,encoding 55 amino acids.In the transcriptional level,the three ATP synthase subunits w.ere widely distributed in some,immune related tissues.In the expression pattern analysis,MjATP5I,MjATP5L and MjATP5E could be upregulated by WSSV challenge in hemocytes and intestine of shrimp.The overexpression assay(injection of recombinant protein)and RNA interference assay showed that MjATP5I and MjATP5L and MjATP5E were beneficial for WSSV replication in M.japonicus.All results demonstrated that MjATP5I,MjATP5L and MjATP5E could promote the WSSV replication in shrimp.The mechanisms still need further study.