Thiamine is a major vitamin that acts as a cofactor in energy metabolism in all organisms, as well as in lipid and amino acid metabolisms, and is associated with many diseases. It is known that glucose starvation decreases the intracellular thiamine pool while increasing oxidative stress tolerance. Earlier, in whole genome analysis, we detected major differences in the expression of genes related to thiamine pathway against oxidative stress in Schizosaccharomyces pombe. We investigated the effects of oxidative stress and glucose repression to thiamine pathway in S. pombe by comparing some genes encoding key enzymes of each related pathway at the transcription level. In the present study, we found that the expression of genes related to thiamine biosynthesis and transport (thi2, thi3, and pho1) increased in wild type and ird11 cells grown in thiamine-rich media under oxidative stress induced by H2O2. Based on our findings, we suggested that there might be an important effect of oxidative stress on thiamine biosynthesis and transport.

Background: Glucose is the preferred carbon and energy source in most organisms and plays an active role in the regulation of many biological processes. However, an excess of glucose leads to such undesirable conditions as diabetes and age-related diseases. Since Schizosaccharomyces pombe homologous of many human genes, it offers several advantages for the investigation of the molecular mechanisms underlying human disease and aging studies. We have identified two glucose-repression-resistant mutants (ird5 and ird11) of S. pombe. Objectives: We aimed to investigate the possible relationship between lifespan extension and oxidative stress response induced by exposure to hydrogen peroxide alongside the trehalose accumulation level by using the two S. pombe mutants (i.e. ird5 and ird11), which are repressed by glucose and are resistant to oxidative stress. Materials and Methods: We employed trehalose accumulation measurement and colony-forming unit (CFU) counting using the ird mutants in exponential and stationary phases and compared them to the wild type grown in repressed, de-repressed, and stressed conditions to clarify the possible relationship between glucose signaling, oxidative stress response, and lifespan in S. pombe. Results: The lifespan of the ird5 mutant was significantly longer that of either the ird11 mutant or the wild type cells. Under repressed condition, the trehalose content was increased remarkably on the 3rd day of the study in the ird11 mutant and the wild type. Under de-repressed condition, the level of intracellular trehalose was notably increased on the 3rd day in ird11. Under stressed condition, the trehalose level in ird11 was increased on the 3rd day as a pattern similar to that observed in the wild type. Conclusions: Our results demonstrated no significant correlation between the ird5 lifespan and the trehalose concentration. Likewise, the correlation between lifespan extension, trehalose accumulation, and cellular resistance to hydrogen peroxide was not significant. Keywords: Trehalose; Glucose; Repression; Fission Yeast

We looked for changes in gene expression and novel genes that could be involved in the interaction between glucose repression and oxidative stress response in the fission yeast, Schizosaccharomyces pombe, using a constitutive invertase mutant, ird11, which is resistant to glucose. BLAST analysis was made of the S. pombe genome database of cDNAs whose expression ratios differentially decreased or increased upon exposure to mild oxidative stress in this mutant compared to the wild type. Genes with this type of activity were identified as rpl302, encoding 60S ribosomal protein L3, and mpg1, encoding mannose-1-phosphate guanyltransferase; their expression patterns were measured using quantitative real-time PCR. We found that the expression levels of rpl302 and mpg1 genes in ird11 under unstressed conditions were increased compared to those of the wild type. Under stress conditions, the expression levels of the rpl302 gene were decreased in both strains, while mpg1 expression levels remained unchanged. These results suggest that these genes play a role in the response to oxidative stress in this mutant strain.

The resistant to glucose repression mutants of Schizosaccharomyces pombe (ird5, ird13, and ird14) have a high tolerance to oxidative stress induced by H2O2. In all ird mutants, the increased expression level of the fbp1 gene can be interpreted as a lack of glucose repression in these mutants. To investigate the mechanisms of the oxidative stress response in ird mutants, we analyzed the transcription of stress response-related genes, sod1, ctt1, atf1, pap1, and sty1, under stressed and non-stressed conditions. We then analyzed the phosphorylation state of the Sty1-MAP kinase in ird mutants. Our findings support the concept of an adaptive response to oxidative stress in these mutants. In addition, these results imply that either glucose signaling mechanisms leading to glucose repression and glucose utilization as an energy source are regulated apart from each other or, like Saccharomyces cerevisiae, S. pombe might have additional glucose detection systems.