Project description:Here we used functional proteomics approaches to identify an upstream transcription complex (USTC) that is required for piRNA biogenesis. The complex contains PRDE-1, SNPC-4, TOFU-4, and TOFU-5, all of which are enriched on the two piRNA clusters on chromosome IV and form distinct piRNA foci in the nucleus.
Project description:Soybean and fish contains various active components that were reported to prevent cardiovasucular diseases. Epidemiological studies indicated that Asian eating patterns, consisting of daily seafood and/or soy consumption, confer protection against cardiovascular diseases. In this context, we expected that simultaneous intake of soybean and fish may be greatly beneficial in a manner different from the ingestion of the individual food. To understand the detailed mechanism for modulating the lipid metabolism by co-ingestion of a soy food (tofu) and fish oil, we investigated the global changes in hepatic mRNA expression in rats by using a microarray analysis. Generally, ingestion of tofu contributed to prevent the mRNA expressions involved in hepatic lipogenesis, whereas fish oil facilitated the mRNA expressions related to the degradation of hepatic fatty acids. In terms of mRNA expression, the interaction of two dietary factors was limited. The expression data was normalized and summarized by using SuperNORM data service (Skylight Biotech Inc.). Significance of expressional change among groups was tested by 2-way ANOVA on the normalized CEL data, which was deposited in a tab-separated ASCII text format. Principal components were identified on the summarized gene data. Rats were divided into four groups each with similar average body weights and assigned experimental diets for 21 days as follows: casein and soybean oil diet (CS); casein and fish oil diet (CF); tofu and soybean oil diet (TS); tofu and fish oil diet (TF). Total protein and fat content of each diet was 200 and 150 g/kg, respectively. Soybean oil diets (CS and TS) contain 150 g/kg diet of soybean-derived fat (i.e. soybean oil and freeze-dried tofu), and 50 g/kg of fish oil was replaced with the same amount of soybean-derived fat for fish oil diets (CF and TF). Animals were allowed free access to food and water.
Project description:Piwi-interacting RNAs (piRNAs) play significant roles in suppressing transposons and nonself nucleic acids, maintaining genome integrity, and defending against viral infections, and are essential for fertility in a variety of organisms. In C. elegans, most piRNA precursors are transcribed by RNA polymerase II in the nucleus and are subjected to a number of processing and maturation steps. However, the biogenesis of piRNAs is still not fully understood. We used functional proteomics to study piRNA biogenesis in C. elegans and identified a piRNA processing and chromosome segregation (PICS) complex. The PICS complex contains two known piRNA biogenesis factors, TOFU-6 and PID-1, and three new proteins PICS-1, TOST-1, and ERH-2, which exhibit dynamic localization among different subcellular compartments. In the germlines of gravid animals, the PICS complex contains TOFU-6/PICS-1/ERH-2/PID-1, is largely concentrated at the perinuclear granule zone and engages in piRNA processing. During early embryogenesis, the TOFU-6/PICS-1/ERH-2/TOST-1 complex accumulates in the nucleus and plays essential roles in chromosome segregation and cell division. Interestingly, the functions of these factors in mediating chromosome segregation are independent of piRNA production. Therefore, we speculate that differential compositions of PICS factors may help cells coordinate distinct cellular processes.
Project description:The effects of freeze-dried tofu, a traditional Japanese soy food, were compared with those of major active soy components, protein and isoflavone, by observing physiological differences and global transcriptomes in the liver of male rats. The GeneChip data was normalized and summarized by using SuperNORM data service (Skylight Biotech Inc.). Significance of expressional change among groups was tested by 2-way ANOVA on the normalized CEL data, which was deposited in a tab-separated ASCII text format. Principal components were identified on the summarized gene data. Rats were randomly divided into 6 groups of 5 samples and assigned experimental diets for 14 days. The experimental diets were as follows: casein diet (C); C containing isoflavone (CI) soy protein diet (S); S containing isoflavone (SI); a diet containing 100 g/kg each of protein derived from casein and freeze-dried tofu (T10); a diet containing 200 g/kg of protein derived from freeze-dried tofu (T20). CI and SI were supplemented with a mixture of isoflavones to match the isoflavone level of T20.
Project description:Piwi-interacting RNAs (piRNAs) are genomically encoded small RNAs that engage Piwi Argonaute proteins to direct mRNA surveillance and transposon silencing. Despite advances in understanding piRNA pathways and functions, how the production of piRNA is regulated remains elusive. Here, using a genetic screen, we identify casein kinase II (CK2) as a factor required for piRNA pathway function. We show that CK2 is required for the localization of PRG-1 and for the proper localization of several factors that comprise the ‘upstream sequence transcription complex’ (USTC), which is required for piRNA transcription. Loss of CK2 impairs piRNA levels suggesting that CK2 promotes USTC function. We identify the USTC component twenty-one-U fouled-up 4 (TOFU-4) as a direct substrate for CK2. Our findings suggest that phosphorylation of TOFU-4 by CK2 promotes the assembly of USTC and piRNA transcription. Notably, during the aging process, CK2 activity declines, resulting in the disassembly of USTC, decreased piRNA production, and defects in piRNA-mediated gene silencing, including transposons silencing. These findings highlight the significance of posttranslational modification in regulating piRNA biogenesis and its implications for the aging process. Overall, our study provides compelling evidence for the involvement of a posttranslational modification mechanism in the regulation of piRNA biogenesis.
2024-07-01 | GSE239315 | GEO
Project description:Metagenomics of stinky tofu brine