Overexpression of a Brix Domain-Containing Ribosome Biogenesis Factor ARPF2 and its Interactor ARRS1 Causes Morphological Changes and Lifespan Extension in Arabidopsis thaliana

The Brix domain is a conserved domain in several proteins involved in ribosome biogenesis in yeast and animals. In the Arabidopsis genome, six Brix domain-containing proteins are encoded; however, their molecular functions have not been fully characterized, as yet. Here we report the functional analysis of a Brix domain-containing protein, ARPF2, which is homologous to yeast Rpf2 that plays an essential role in ribosome biogenesis as a component of the 5S ribonucleoprotein particle. By phenotypic characterization of arpf2 mutants, histochemical GUS staining, and analysis using green fluorescence protein, we show that ARPF2 is an essential and ubiquitously expressed gene encoding a nucleolar protein. Co-immunoprecipitation and split-GFP-based bimolecular fluorescence complementation assays revealed that ARPF2 interacts with a protein named ARRS1, which is homologous to yeast Rrs1 that forms a complex with Rpf2 in yeast. Furthermore, the result of RNA immunoprecipitation assay indicated that ARPF2 interacts with 5S ribosomal RNA (rRNA) or the precursor of 5S rRNA, as well as with the internal transcribed spacer 2 in the precursors of 25S rRNA. Most intriguingly, we found that the overexpression of ARPF2 and ARRS1 leads to characteristic phenotypes, including short stem, abnormal leaf morphology, and long lifespan, in Arabidopsis. These results suggest that the function of Brix domain-containing ARPF2 protein in ribosome biogenesis is intimately associated with the growth and development in plants

The lectin‐like domain of thrombomodulin is a drug candidate for both prophylaxis and treatment of liver ischemia and reperfusion injury in mice

Ischemia and reperfusion injury (IRI) can occur in any tissue or organ. With respect to liver transplantation, the liver grafts from donors by definition experience transient ischemia and subsequent blood reflow. IRI is a problem not only in organ transplantation but also in cases of thrombosis or circulatory disorders such as mesenteric ischemia, myocardial, or cerebral infarction. We have reported that recombinant human soluble thrombomodulin (rTM), which is currently used in Japan to treat disseminated intravascular coagulation (DIC), has a protective effect and suppresses liver IRI in mice. However, rTM may not be fully safe to use in humans because of its inherent anticoagulant activity. In the present study, we used a mouse liver IRI model to explore the possibility that the isolated lectin-like domain of rTM (rTMD1), which has no anticoagulant activity, could be effective as a therapeutic modality for IRI. Our results indicated that rTMD1 could suppress ischemia and reperfusion-induced liver damage in a dose-dependent manner without concern of associated hemorrhage. Surprisingly, rTMD1 suppressed the liver damage even after IR insult had occurred. Taken together, we conclude that rTMD1 may be a candidate drug for prevention of and therapy for human liver IRI without the possible risk of hemorrhage

Stability and safety estimates and tests of a superconducting bus-line for large-scale superconducting coils

We have been developing a flexible superconducting bus-line as a unit electrical feeder between large-scale superconducting coils and their power supplies away from the coils. The designed superconducting bus-line consists of a pair of +/- aluminum stabilized NbTi/Cu compacted strand cables and a coaxial four-channel transfer line. A full-scale model of the SC bus-line (20 m long) has been constructed and tested successfully up to 40 kA without a quench under the short-circuit condition. Stability tests were also done by inducing a forced quench with heaters. A minimum propagation current larger than 32.5 kA was confirmed. Thus, the bus-line was cryogenically stabilized at the rated current of 30 kA. We have examined the test results and evaluated the stability and safety margins of this bus-line. The design criteria for a superconducting bus-line are also shown for large-scale superconducting coils with operating current as a parameter

Development and tests of a flexible superconducting bus-line for the Large Helical Device

A flexible superconducting bus-line is proposed as an electrical feeder between the superconducting coils of the Large Helical Device (LHD) and the device\u27s power supplies. The bus-line consists of superconducting cables and a cryogenic flexible transfer-line. A specially developed aluminum stabilized NbTi/Cu compacted strand cable satisfies requirements for large current capacity, high stability, high reliability and flexibility. A full-scale model with a length of 20 m was designed and constructed to investigate the feasibility and performance of the superconducting bus-line. Its fabrication, transportation, installation, cooling and excitation tests were successfully carried out. The bus-line was very stable and could be excited up to 40 kA (rated current is 30 kA) without a quench. The stability, current distribution and heat load were also measure

Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system

Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins

カスパーゼ3(様)蛋白分解酵素活性のレドックス制御機構、チオレドキシンとシトクロムcによる制御機構に関する研究

京都大学0048新制・課程博士博士(医学)甲第8241号医博第2197号新制||医||731(附属図書館)UT51-2000-F145京都大学大学院医学研究科外科系専攻(主査)教授 日合 弘, 教授 伊藤 和彦, 教授 山岡 義生学位規則第4条第1項該当Doctor of Medical ScienceKyoto UniversityDA

Proteasome-dependent Degradation of Cyclin D1 in 1-Methyl-4-phenylpyridinium Ion (MPP+)-induced Cell Cycle Arrest

1-Methyl-4-phenylpyridinium ion (MPP+), an active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, induces cell death and inhibition of cell proliferation in various cells. However, the mechanism whereby MPP+ inhibits cell proliferation is still unclear. In this study, we found that MPP+ suppressed the proliferation with accumulation in G1 phase without inducing cell death in p53-deficient MG63 osteosarcoma cells. MPP+ induced hypophosphorylation of retinoblastoma protein and rapidly down-regulated the protein but not mRNA levels of cyclin D1 in MG63 cells. The down-regulation of cyclin D1 protein was suppressed by a proteasome inhibitor, MG132. The cyclin D1 down-regulation by MPP+ was also observed in p53-positive PC12, HeLa S3, and HeLa 0 cells, which are a subclone of HeLa S3 lacking mitochondrial DNA. Moreover, MPP+ dephosphorylated Akt in PC12 cells, which was rescued by the pretreatment with nerve growth factor. In addition, the pretreatment with nerve growth factor or lithium chloride, a glycogen synthase kinase-3 inhibitor, suppressed the cyclin D1 down-regulation caused by MPP+. Our results demonstrate that MPP+ induces cell cycle arrest independently of its mitochondrial toxicity or the p53 status of the target cells, but rather through the proteasome- and phosphatidylinositol 3-Akt-glycogen synthase kinase-3-dependent cyclin D1 degradation