Study On The Expression Of Trz2 Gene In Schizosaccharomyces Pombe And The Influence Of Mtf2 Gene On Mitochondrial Genome Expression

In eukaryotes,mitochondrial genome co-operated with nuclear genome in biological and synthetic aspects.The different nuclear gene protein was involved in the mitochondrial gene of transcription,RNA processing,and translation.S.pombe have two mitochondrial proteins Trz2 and Mtf2,which were encoded the nuclear genes.And Trz2 and Mtf2 play the important roles in the mitochondria.We have demonstrated that Trz2 was involved in mitochondrial tRNA precursor 3'-end processing and the maturation of the 5'-end of mitochondrial mRNAs and rRNAs in S.pombe.We have also demonstrated that overxpression of SpTrz2p affected the cell cycle.In addition,overxpression of SpTrz2p resulted in the lethal cells.In our study,the growth of yHDl containing a temperature-sensitive allele of trz2 was defected in the non-fermentation medium.We realized that SpTrz2p was associated with mitochondrial function.SpTrz2 is low expression protein in anti-SpTrz2p antibody and anti-SpTrz2p-N-terminal assay.5' RACE was performed to map the trz2 transcription start site located 60 bp upstream from the initiation codon in S.pombe wild cells.The?-galactosidase assay was used for the study of the core trz2 promoter regions.We found that the repressive region(-111 to-54),also called the URS.And overexpression of rrn7 up-regulated HomolD box-directed transcription of trz2 gene.In addition,HomolD box was evolutionary conservation for each of all four trz2 genes regulation in the microcommunity of fission yeast on the bioinformatic analysis.Therefore,we showed that the trz2 expression was regulated by a weak TATA box,a repressive region and a positive HomolD box at the critical balance.S.cerevisiae Mtf2 homologous protein involved in mitochondrial RNA processing and translation.S.pombe mitochondrial protein Mtf2 is an unknown protein.We studyed the cellular morphology of ?mtf2 strain and the determination of ?mtf2 cell growth curve to understand the growth characteristics of ?mtf2 strain.The growth of?mtf2 strain was defected in the non-fermentation medium.We considered that S.pombe Mtf2 was associated with mitochondrial function.By using Real-Time PCR,in vivo 35S labeling of ?mtf2 mutant and Western-Blot,we see that mRNA level and protein level of ?mtf2 strain are affected in the mitochondrial genome.Experiments show that S.pombe mtf2 gene influenced the expression of the mitochondrial genome.