The deletion of ppr2 interferes iron sensing and leads to oxidative stress response in Schizosaccharomyces pombe.

Pentatricopeptide repeat proteins are involved in mitochondrial both transcriptional and posttranscriptional regulation. Schizosaccharomyces pombe Ppr2 is a general mitochondrial translation factor that plays a critical role in the synthesis of all mitochondrial DNA-encoded oxidative phosphorylation subunits, which are essential for mitochondrial respiration. Our previous analysis showed that ppr2 deletion resulted in increased expression of iron uptake genes and caused ferroptosis-like cell death in S. pombe. In the present work, we showed that deletion of ppr2 reduced viability on glycerol- and galactose-containing media.Php4 is a transcription repressor that regulates iron homeostasis in fission yeast. We found that in the ppr2 deletion strain, Php4 was constitutively active and accumulated in the nucleus in the stationary phase. We also found that deletion of ppr2 decreased the ferroptosis-related protein Gpx1 in the mitochondria. Overexpression of Gpx1 improves the viability of Δppr2 cells. We showed that the deletion of ppr2 increased the production of ROS, downregulated heme synthesis and iron-sulfur cluster proteins, and induced stress proteins. Finally, we observed the nuclear accumulation of Pap1-GFP and Sty1-GFP, suggesting that Sty1 and Pap1 in response to cellular stress in the ppr2 deletion strain. These results suggest thatppr2 deletion may cause mitochondrial dysfunction, which is likely to lead to iron-sensing defect and iron starvation response, resulting in perturbation of iron homeostasis and increased hydroxyl radical production. The increased hydroxyl radical production triggers cellular responses in theppr2 deletion strain.

Learning multi-modal brain tumor segmentation from privileged semi-paired MRI images with curriculum disentanglement learning.

Since the brain is the human body's primary command and control center, brain cancer is one of the most dangerous cancers. Automatic segmentation of brain tumors from multi-modal images is important in diagnosis and treatment. Due to the difficulties in obtaining multi-modal paired images in clinical practice, recent studies segment brain tumors solely relying on unpaired images and discarding the available paired images. Although these models solve the dependence on paired images, they cannot fully exploit the complementary information from different modalities, resulting in low unimodal segmentation accuracy. Hence, this work studies the unimodal segmentation with privileged semi-paired images, i.e., limited paired images are introduced to the training phase. Specifically, we present a novel two-step (intra-modality and inter-modality) curriculum disentanglement learning framework. The modality-specific style codes describe the attenuation of tissue features and image contrast, and modality-invariant content codes contain anatomical and functional information extracted from the input images. Besides, we address the problem of unthorough decoupling by introducing constraints on the style and content spaces. Experiments on the BraTS2020 dataset highlight that our model outperforms the competing models on unimodal segmentation, achieving average dice scores of 82.91%, 72.62%, and 54.80% for WT (the whole tumor), TC (the tumor core), and ET (the enhancing tumor), respectively. Finally, we further evaluate our model's variable multi-modal brain tumor segmentation performance by introducing a fusion block (TFusion). The experimental results reveal that our model achieves the best WT segmentation performance for all 15 possible modality combinations with 87.31% average accuracy. In summary, we propose a curriculum disentanglement learning framework for unimodal segmentation with privileged semi-paired images. Moreover, the benefits of the improved unimodal segmentation extend to variable multi-modal segmentation, demonstrating that improving the unimodal segmentation performance is significant for brain tumor segmentation with missing modalities. Our code is available at https://github.com/scut-cszcl/SpBTS.

Mrz1, a Novel Mitochondrial Outer Membrane RING Finger Protein, is Degraded Through the Ubiquitin-Proteasome Pathway in Schizosaccharomyces pombe.

The RING (Really Interesting New Gene) finger proteins are a large diverse group of Zinc finger proteins. Many determined RING finger proteins are ubiquitin-protein E3 ligases and RING E3s are the most abundant type of ubiquitin ligase. RING finger and RING finger E3s have been discovered in many organisms where they play various functions, including DNA repair, ubiquitination and mitochondrial protein quality control. In this study, we identified a novel mitochondrial protein (SPBC16G5.03) with predicted RING finger domain within an N-terminal 21-60 amino acids and named it Mrz1 (mitochondrial RING finger protein). Our results showed that Mrz1 is localized in the mitochondrial outer membrane. Deletion of mrz1 did not affect cell growth in an unstressed state, but increases sensitivity to selenite. We showed that Mrz1 was degraded during the stationary phase and blocked by addition proteasome inhibitor MG132. We further showed that the E2 enzyme Ubc13 was identified among 8 candidate proteins as the ubiquitin-conjugating enzyme in this system. These data suggested that the Mrz1 was degraded likely through the ubiquitin-proteasome system.

Loss of PPR protein Ppr2 induces ferroptosis-like cell death in Schizosaccharomyces pombe.

Ferroptosis is a form of iron- and lipid peroxidation-mediated programmed cell death that occurs widely in mammalian cells. However, this phenomenon is rarely reported in unicellular eukaryotes. Here, we address whether ferroptosis occurs in the model unicellular eukaryote Schizosaccharomyces pombe (S. pombe). Deletion of the pentatricopeptide repeat (PPR) gene ppr2 encoding as a general mitochondrial translation factor required for mitochondrial translation disrupts iron homeostasis and induces oxidative stress, resulting in loss of cell viability. The small-molecular ferroptosis inhibitors deferoxamine (DFO) and ferrostatin-1 (Fer-1) partially rescued the ppr2 deletion-induced cell death. The amount of malondialdehyde, a lipid peroxidation marker, in Δppr2 cells was higher than that in wild type. Using C11-BODIPY 581/591, an oxidation-sensitive fluorescent lipid peroxidation probe, we showed that Δppr2 cells have a large amount of lipid peroxidation compared to wild-type cells. Deletion of ferric reductase transmembrane component 1 (frp1) encoding S. pombe ferric reductase, which is required for ferric iron uptake, partially rescued the cell death of Δppr2 cells. Our results suggest that ppr2 deletion causes an imbalance in iron homeostasis and redox, leading to ferroptosis-like cell death in S. pombe.

Schizosaccharomyces pombe Ppr10 is required for mitochondrial translation.

The mitochondrial genome encodes key components of the oxidative phosphorylation (OXPHOS) system, whose expression is essential for mitochondrial functions. We have previously shown that deletion of the Schizosaccharomyces pombe ppr10 encoding a pentatricopeptide repeat protein severely reduces the mature levels of intron-containing mitochondrial transcripts cox1 and cob1, and severely impairs mitochondrial translation. In this study, we examined the possibility that the reduced levels of Cox1 and Cob1 proteins in cells were due to lowered levels of cox1 and cob1 mRNAs. We found that deletion of ppr10 did not affect the levels of mature cox1 and cob1 mRNAs in a mitochondrial intronless background. However, synthesis of Cox1 and Cob1 proteins were still severely affected by deletion of ppr10 in a mitochondrial intronless background. Consistent with this, we found that deletion of mitochondrial introns could not rescue the respiratory growth defect of Δppr10 cells. Our results reveal that Ppr10 is not required for the stability of cox1 and cob1 mRNAs, and provide further support for the idea that Ppr10 plays a critical role in mitochondrial translation.

Experimental Study on Vibration Characteristics of Unit-Plate Ballastless Track Systems Laid on Long-Span Bridges Using Full-Scale Test Rigs.

In this work, we present a series of hammering tests on full-scale unit-plate ballastless tracks used for long-span bridges. There is no denying that it is a new attempt to pave ballastless tracks on high-speed railway long-span bridges; the related issues deserve to be studied, and especially the vibration characteristics. Hence, the vibration characteristics and transmission rules of the ballastless track with geotextile or rubber isolation layers are explored, and the vibration reduction effect of the rubber isolation layer is analyzed. The main conclusions are as follows: the isolation layers change vibration modes and transmission characteristics of ballastless tracks; the introduction of the rubber isolation layer makes the excited vibration frequency range of the ballastless track concentrated; and the vibrations of the ballastless track with the rubber isolation layers are stable. Moreover, the rubber isolation layer has an obvious attenuation effect on vibration transmission in ballastless track structures. When the vibration is transmitted from the rail to the bridge deck, the vibration level differences of the ballastless track with rubber isolation layers are 20 dB larger than that of the ballastless track with the geotextile isolation layers. The vibration attenuation rate of the rubber isolation layer is about ten times larger than that of geotextile isolation layer.

Schizosaccharomyces pombe Mti2 and Mti3 act in conjunction during mitochondrial translation initiation.

Mitochondrial DNA encodes key subunits of the oxidative phosphorylation complexes essential for ATP production. Translation initiation in mitochondria requires two general factors, mtIF2 and mtIF3, whose counterparts in bacteria are essential for protein synthesis. In this study, we report the characterization of the fission yeast Schizosaccharomyces pombe mtIF2 (Mti2) and mtIF3 (Mti3). Deletion of mti2 impairs cell growth on the respiratory medium. The growth defect of the mti2 deletion mutant can be suppressed by expressing IFM1, the Saccharomyces cerevisiae homolog of Mti2, demonstrating functional conservation between the two proteins. Deletion of mti2 also impairs mitochondrial protein synthesis. Unlike mti2, deletion of mti3 does not affect cell growth on respiratory media and mitochondrial translation. However, deletion of mti3 exacerbates the growth defect of the Δmti2 mutant, suggesting that the two proteins have distinct, but partially overlapping functions during the process of mitochondrial translation initiation in S. pombe. Both Mti2 and Mti3 are associated with the small subunit of the mitochondrial ribosome (mitoribosome). Disruption of mti2, but not mti3, causes dissociation of the mitoribosome and also abolishes Mti3 binding to the small subunit of the mitoribosome. Our results suggest that Mti2 and Mti3 bind in a sequential manner to the small subunit of the mitoribosome and that Mti3 facilitates the function of Mti2 in mitochondrial translation initiation. Our findings also support the view that the importance of the mitochondrial translation initiation factors varies among the organisms.

Overexpression of Schizosaccharomyces pombe tRNA 3'-end processing enzyme Trz2 leads to an increased cellular iron level and apoptotic cell death.

The fission yeast Schizosaccharomyces pombe has two tRNase ZL genes (trz1 and trz2) involved in nuclear and mitochondrial tRNA 3'-end processing, respectively. Overexpression of trz2 but not trz1 is toxic to cells. In the present work, we showed that trz2 overexpression led to apoptotic cell death, as revealed by DAPI and Annexin V-FITC staining. Overexpression of trz2 also caused a loss of mitochondrial membrane potential and an increased reactive oxygen species (ROS) formation. These effects required mitochondrial localization but not its catalytic activity. RNA sequencing (RNA-seq) analysis revealed increased expression levels of genes involved in iron uptake and/or iron homeostasis, suggesting an elevated level of intracellular iron in the trz2-overexpressing cells. Indeed, we showed that overexpressing trz2 increased the level of intracellular iron by ∼2-fold. We further showed that the iron chelator, bathophenanthroline disulfonic acid (BPS) nearly restored the viability of trz2-overexpression cells and reduced ROS levels in the cells. These results suggest that trz2 overexpression may cause mitochondrial dysfunction, which is likely to lead to perturbation of iron homeostasis, ROS accumulation and induction of apoptotic cell death in S. pombe.