Functional Study On Trz2 Involved In Schistosoma Japonicum In Mitochondrial RNA Processing And Apoptosis

Mitochondria are essential organelles that harbor a reduced genome. Recent studies implicate metabolism defects of mitochondrial result in the pathophysiology of several human diseases, including AIDS, neurodegenerative disease, and cancer. S. pombe is currently used as model organisms for mitochondrial genetics and molecular biology and have provided a wealth of data on various aspects of organellar biology and the nucleo-mitochondrial interactions.In this study, the mitochondrial of S. pombe was purified and RNAs including tRNAs and small RNAs were harvested from isolated mitochondria, and the quantity and purity of the harvested RNAs were validated by electrophoresis and qRT-PCR. We used high-throughput RNA sequencing and traditional RNA biology techniques to study the mitochondrial transcriptome of S. pombe. Sequencing of the mitochondrial transcriptome revealed a description of RNAs boundaries and wide variation in gene expression that indicates the existence of complex posttranscriptional regulatory mechanisms. We determined the cleavage sites of rnl, atp6 and tRNAsn, and to verify the reliability of the RNA-seq results, CR-RT-PCR analyses was performed. trz2, a tRNase ZL gene in Schizosaccharomyces pombe, has high homologe to the ELAC2 as a gene therapy a prosate cancer-associated protein. Trz2 is targeted to mitochondria and is involved in mitochondrial tRNA 3’end processing. Here, we have global analyzed mitochondrial RNAs processing and mitochondrial gene expression by inactivation of Trz2. Our data have shown that Trz2 is responsible for 3’ end processing of mitochondrial tRNAs and inactivation of Trz2 affects the maturations of the 5’-end of mitochondrial mRNAs and rRNAs.Mitochondria have been shown to play key roles in many instances of apoptosis. Here, we report that overexpression of Trz2 in Schizosaccharomyces pombe induces cell death with apoptosis markers. Nuclear condensation and the fragmentation of chromatin were observed by DAPI stain. About 3 flods of cells overexpressing of Trz2 were stained with AnnexinV-FITC compared with the control cells. And apoptosis-inducing factor(aif1) upregulated by RT-PCR. Further fluorescence microscopy revealed OETrz2 had fragmented mitochondrial with mitoTracker and showed an intense intracellular staining by DHR. Apoptotic death caused by overexpression of Trz2 required its mitochondria target domain. There is no report about Trz2 function and apoptosis related protein in S. pombe.Most of proteins have same functions in S. pombe cells and mammalian cells, and it provides a good model system of molecular genetics studies of eukaryotic genes. We have used fission yeast as a model system to study the mitochondrial RNA processing and the mechanism of apoptosis helps to understand the occurrence and mechanism of RNA processing and apoptosis related diseases for disease prevention and treatment of the new strategy.