ER Stress Proteins in Autoimmune and Inflammatory Diseases

Over the past two decades, heat shock proteins (HSPs) have been implicated in inflammatory responses and autoimmunity. HSPs were originally believed to maintain protein quality control in the cytosol. However, they also exist extracellularly and appear to act as inflammatory factors. Recently, a growing body of evidence suggested that the other class of stress proteins such as, endoplasmic reticulum (ER) stress proteins, which originally act as protein quality control factors in the secretory pathway and are induced by ER stress in inflammatory lesions, also participate in inflammation and autoimmunity. The immunoglobulin heavy-chain binding protein (Bip)/glucose-regulated protein 78 (GRP78), calnexin, calreticulin, glucose-regulated protein 94 (GRP94)/gp96, oxygen regulated protein 150 (ORP150)/glucose-regulated protein 170 (GRP170), homocysteine-induced ER protein (Herp) and heat shock protein 47 (hsp47)/Serpin H1, which are expressed not only in the ER but also occasionally at the cell surface play pathophysiological roles in autoimmune and inflammatory diseases as pro- or anti-inflammatory factors. Here we describe the accumulating evidence of the participation of ER stress proteins in autoimmunity and inflammation and discuss the critical differences between the two classes of stress proteins

Gp78 Cooperates with RMA1 in Endoplasmic Reticulum-associated Degradation of CFTR F508

Misfolded or improperly assembled proteins in the endoplasmic reticulum (ER) are exported into the cytosol and degraded via the ubiquitin–proteasome pathway, a process termed ER-associated degradation (ERAD). Saccharomyces cerevisiae Hrd1p/Der3p is an ER membrane-spanning ubiquitin ligase that participates in ERAD of the cystic fibrosis transmembrane conductance regulator (CFTR) when CFTR is exogenously expressed in yeast cells. Two mammalian orthologues of yeast Hrd1p/Der3p, gp78 and HRD1, have been reported. Here, we demonstrate that gp78, but not HRD1, participates in ERAD of the CFTR mutant CFTRΔF508, by specifically promoting ubiquitylation of CFTRΔF508. Domain swapping experiments and deletion analysis revealed that gp78 binds to CFTRΔF508 through its ubiquitin binding region, the so-called coupling of ubiquitin to ER degradation (CUE) domain. Gp78 polyubiquitylated in vitro an N-terminal ubiquitin-glutathione-S-transferase (GST)-fusion protein, but not GST alone. This suggests that gp78 recognizes the ubiquitin that is already conjugated to CFTRΔF508 and catalyzes further polyubiquitylation of CFTRΔF508 in a manner similar to that of a multiubiquitin chain assembly factor (E4). Furthermore, we revealed by small interfering RNA methods that the ubiquitin ligase RMA1 functioned as an E3 enzyme upstream of gp78. Our data demonstrates that gp78 cooperates with RMA1 with E4-like activity in the ERAD of CFTRΔF508

Identification of RNF213 as a Susceptibility Gene for Moyamoya Disease and Its Possible Role in Vascular Development

もやもや病感受性遺伝子の特定とその機能についての発見. 京都大学プレスリリース. 2011-7-21.Background Moyamoya disease is an idiopathic vascular disorder of intracranial arteries. Its susceptibility locus has been mapped to 17q25.3 in Japanese families, but the susceptibility gene is unknown. Methodology/Principal Findings Genome-wide linkage analysis in eight three-generation families with moyamoya disease revealed linkage to 17q25.3 (P<10-4). Fine mapping demonstrated a 1.5-Mb disease locus bounded by D17S1806 and rs2280147. We conducted exome analysis of the eight index cases in these families, with results filtered through Ng criteria. There was a variant of p.N321S in PCMTD1 and p.R4810K in RNF213 in the 1.5-Mb locus of the eight index cases. The p.N321S variant in PCMTD1 could not be confirmed by the Sanger method. Sequencing RNF213 in 42 index cases confirmed p.R4810K and revealed it to be the only unregistered variant. Genotyping 39 SNPs around RNF213 revealed a founder haplotype transmitted in 42 families. Sequencing the 260-kb region covering the founder haplotype in one index case did not show any coding variants except p.R4810K. A case-control study demonstrated strong association of p.R4810K with moyamoya disease in East Asian populations (251 cases and 707 controls) with an odds ratio of 111.8 (P = 10−119). Sequencing of RNF213 in East Asian cases revealed additional novel variants: p.D4863N, p.E4950D, p.A5021V, p.D5160E, and p.E5176G. Among Caucasian cases, variants p.N3962D, p.D4013N, p.R4062Q and p.P4608S were identified. RNF213 encodes a 591-kDa cytosolic protein that possesses two functional domains: a Walker motif and a RING finger domain. These exhibit ATPase and ubiquitin ligase activities. Although the mutant alleles (p.R4810K or p.D4013N in the RING domain) did not affect transcription levels or ubiquitination activity, knockdown of RNF213 in zebrafish caused irregular wall formation in trunk arteries and abnormal sprouting vessels. Conclusions/Significance We provide evidence suggesting, for the first time, the involvement of RNF213 in genetic susceptibility to moyamoya disease

Space charge behavior in multi-layer oil-paper insulation under different DC voltages and temperatures

Oil-paper as a reliable insulation system is widely used in power transformers and cables. The dielectric properties of oil-paper insulation play an important role in the reliable operation of power equipment. However, the formation and dynamics of space charge can affect the performance of insulation material. In this paper, space charge in oil-paper insulation system has been investigated using the pulsed electroacoustic (PEA) technique. A series of measurements were carried out when the insulation system was subjected to different applied voltages at different temperatures. Charge behavior in the insulation system has been analyzed and the influence of temperature on charge dynamics was discussed. The test results show that homocharge injection takes place under all the test conditions, the applied dc voltage mainly effect the amount of space charge, while the temperature has greater influence on the distribution and mobility of space charge inside oil-paper sample

Creatine Supplementation Alleviates Fatigue after Exercise through Anti-Inflammatory Action in Skeletal Muscle and Brain

Eccentric exercise induces muscle damage and inflammation, resulting in a state of reduced physical activity with muscle dysfunction and a feeling of tiredness after exercise. Creatine is known to act as an energy buffer, but it has also been suggested to exert inhibitory effects on muscle damage and peripheral inflammation. The purpose of this study was to test the hypothesis that creatine supplementation alleviates fatigue after eccentric exercise and to explore the mechanism of this effect. C57BL/6J mice were fed an AIN-93G-formulated control diet or a creatine-containing diet for 6 days and were then subjected to downhill running, a model of eccentric exercise, to assess the effects on the total creatine concentrations in skeletal muscle and brain tissue, spontaneous activity, the urine concentration of titin N-fragment, and inflammatory gene expression. The results showed that creatine supplementation significantly increased the total creatine concentrations in skeletal muscle and brain tissue. Furthermore, spontaneous activity significantly decreased after downhill running and creatine supplementation maintained a significantly higher level of spontaneous activity. In addition, creatine supplementation significantly suppressed the downhill-running-induced increase in the mRNA expression of genes encoding ICAM-1, E-selectin, CD18, and BKB1R in the soleus muscle and IL-1β in the hypothalamus. On the other hand, creatine supplementation did not clearly influence the urine concentration of titin N-fragment. These results indicate that creatine supplementation may alleviate fatigue after eccentric exercise by partially suppressing inflammation in slow-twitch skeletal muscle and brain tissue