R354X Mutation in PRPF31 Affects Its Capacity in Inflammation Response and Splicing

Pre-mRNA processing factor 31(PRPF31) is a key component of RNA splicing and also a disease-causing gene of Retinitis Pigmentosa (RP). Previously, we found that the nonsense mutation R354X in PRPF31induces RP in a Chinese RP family. In order to investigate the underlining molecular mechanisms of RP pathogenesis induced by this mutation, we generated cell lines stably expressing R354X mutant, wild type (WT) of PRPF31 and corresponding empty vector using HEK293T cells ,the resulting cell lines were used for Long non-coding RNA sequencing(LncRNA -sequencing). The results of LncRNA sequencing showed that, comparing to WT, R354X mutation changed the expression and splicing of coding and non-coding transcripts. Interestingly, in HEK293T and APRE-19 cells, inflammation-associated genes such as IFI6, OAS3and STAT3, enhanced their expression in response to the overexpression of WT PRPF31; however, in R354X mutation cells, those genes’ expression remained basal levels. Moreover, increased H2AFX expression and attenuated growth capacity were found in cells expressing R354X PRPF31. In contrast with WT, R354X mutant showed varied splicing model for dihydrofolate reductase (DHFR) in HEK293T , reduced CPSF1 and SORBS1 mRNAs binding in ARPE-19 ,and its binding with CTNNBL1 was also interfered in ARPE-19 cells. On the other hand, the R354X mutant also increased the level of transcripts read-through. Taken together, R354X mutation in PRPF31 affected cell survival, changed gene’s expression and splicing. Those findings indicate that inflammation and oxidation might contribute the pathogenesis of RP induced by the R354X mutation.

Modification of carbohydrates of food raw materials in the process of thermoplastic extrusion (review)

Extrusion can be considered not only as an effective technology for processing agricultural raw materials into feed and food products, but also as a thermo-mechanical method for modification of the chemical properties of biopolymers. Carbohydrates are the most represented class of organic compounds in raw materials processed by the agro-industrial complex. The assessment of the influence of the processing factor on the final physicochemical and technological properties of various types of carbohydrates included in the chemical composition of raw materials or used as mono-ingredients is an actual task for the food industry. The review considers the issues of extrusion modification of starch in terms of the difference in the properties of amylose and amylopectin as well as the presence of lipids and organic acids in the reaction system. Processes of macromolecular degradation, gelatinization, esterification and the formation of new chemical bonds in dependence on the conditions of extrusion and the composition of mixtures are discussed. The results of studies of the influence of extrusion cooking on the changes in the physicochemical properties of non-starchy polysaccharides, cellulose, araboxylans, inulin, pectin, chitosan, and gums of various origins are presented. It has been shown that extrusion and varying of its operating regimes can significantly affect the nutritional value of extrudates including changing the glycemic index, inactivating antinutritional factors, or increasing their content in extrudates.

CFIm25 regulates human stem cell function independently of its role in mRNA alternative polyadenylation

It has recently been shown that CFIm25, a canonical mRNA 3’ processing factor, could play a variety of physiological roles through its molecular function in the regulation of mRNA alternative polyadenylation (APA). Here, we used CRISPR/Cas9-mediated gene editing approach in human embryonic stem cells (hESCs) for CFIm25, and obtained three gene knockdown/mutant cell lines. CFIm25 gene editing resulted in higher proliferation rate and impaired differentiation potential for hESCs, with these effects likely to be directly regulated by the target genes, including the pluripotency factor rex1. Mechanistically, we unexpected found that perturbation in CFIm25 gene expression did not significantly affect cellular mRNA 3’ processing efficiency and APA profile. Rather, we provided evidences that CFIm25 may impact RNA polymerase II (RNAPII) occupancy at the body of transcribed genes, and promote the expression level of a group of transcripts associated with cellular proliferation and/or differentiation. Further study indicated that CFIm25 association with LEO1, an RNAPII associated factor, might contribute to the effect. Taken together, these results reveal novel mechanisms underlying CFIm25’s modulation in determination of cell fate, and provide evidence that the process of mammalian gene transcription may be regulated by an mRNA 3’ processing factor.

LENG8 regulation of mRNA processing is responsible for the control of mitochondrial activity

The processing of mRNA is essential for the maintenance of cellular and tissue homeostasis. However, the precise regulation of this process in mammalian cells, remains largely unknown. Here we have found that LENG8 represents the mammalian orthologue of the yeast mRNA processing factor Thp3 and Sac3. We go on to demonstrate that LENG8 binds to mRNAs, associates with components of mRNA processing machinery (the TREX complex) and contributes to mRNA nuclear export to the cytoplasm. Loss of LENG8 , leads to aberrant accumulation of poly (A) + RNA in the nucleus, in both Hela cells and murine fibroblasts. Furthermore, the precipitation of LENG8, is associated with an enrichment of both mRNAs and lncRNAs, and approximately half of these are also bound by the TREX component, THOC1. However, LENG8 preferentially binds mRNAs encoding for mitochondrial proteins and depletion of this processing factor, causes a dramatic breakdown in mitochondrial ultrastructure and a reduction in mitochondrial respiratory activity. Conditional deletion of Leng8 in mouse adipose tissues lead to a decreased body weight, and increased adipose thermogenesis. Our work has found an evolutionarily conserved mRNA processing factor that can control mitochondrial activity.

Design of System Level Architecture of Multi-Core with Array Mapping Based on Floating Processing Factor-Fast Fourier Transform for Health Informatics Application

In FFT algorithms memory access patterns prevent multiple architectures from achieving high machine use, particularly when parallel processing is needed to achieve the desired efficiency rates. Beginning with the extremely powerful FFT heart, the on-chip memory hierarchy for the multicored FFT processor, is co-designed and linked on-chip. We have shown that the Floating Processing Factor (FPPE) proposed achieves greater operating rate and lower power for the application of health informatics. This test mechanism aids in omission of faulty cores and autonomous detection also makes elegant multi-core architecture degradation feasible. Experimental results illustrate that the anticipated design is scalable widely in terms of performance overhead and hardware overhead which makes it appropriate to many-cores with more than a thousand processing cores through Low Power and High Speed.

LENG8 regulation of mRNA processing, is responsible for the control of mitochondrial activity

The processing of mRNA is essential for the maintenance of cellular and tissue homeostasis. However, the precise regulation of this process in mammalian cells, remains largely unknown. Here we have found that LENG8 represents the mammalian orthologue of the yeast mRNA processing factor Thp3 and Sac3. We go on to demonstrate that LENG8 binds to mRNAs, associates with components of mRNA processing machinery (the TREX complex) and contributes to mRNA nuclear export to the cytoplasm. Loss of LENG8, leads to aberrant accumulation of poly (A)+ RNA in the nucleus, in both Hela cells and murine fibroblasts. Furthermore, the precipitation of LENG8, is associated with an enrichment of both mRNAs and lncRNAs, and approximately half of these are also bound by the TREX component, THOC1. However, LENG8 preferentially binds mRNAs encoding for mitochondrial proteins and depletion of this processing factor, causes a dramatic breakdown in mitochondrial ultrastructure and a reduction in mitochondrial respiratory activity. Conditional deletion of Leng8 in mouse adipose tissues lead to a decreased body weight, and increased adipose thermogenesis. Our work has found an evolutionarily conserved mRNA processing factor that can control mitochondrial activity.

The Wnt-specific astacin proteinase HAS-7 restricts head organizer formation in Hydra

Background The Hydra head organizer acts as a signaling center that initiates and maintains the primary body axis in steady state polyps and during budding or regeneration. Wnt/beta-Catenin signaling functions as a primary cue controlling this process, but how Wnt ligand activity is locally restricted at the protein level is poorly understood. Here we report a proteomic analysis of Hydra head tissue leading to the identification of an astacin family proteinase as a Wnt processing factor. Results Hydra astacin-7 (HAS-7) is expressed from gland cells as an apical-distal gradient in the body column, peaking close beneath the tentacle zone. HAS-7 siRNA knockdown abrogates HyWnt3 proteolysis in the head tissue and induces a robust double axis phenotype, which is rescued by simultaneous HyWnt3 knockdown. Accordingly, double axes are also observed in conditions of increased Wnt activity as in transgenic actin::HyWnt3 and HyDkk1/2/4 siRNA treated animals. HyWnt3-induced double axes in Xenopus embryos could be rescued by coinjection of HAS-7 mRNA. Mathematical modelling combined with experimental promotor analysis indicate an indirect regulation of HAS-7 by beta-Catenin, expanding the classical Turing-type activator-inhibitor model. Conclusions We show the astacin family protease HAS-7 maintains a single head organizer through proteolysis of HyWnt3. Our data suggest a negative regulatory function of Wnt processing astacin proteinases in the global patterning of the oral-aboral axis in Hydra.