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( Ae Ran Lee ), ( Gyoung Min Kim ), ( Gun Hee Kwon ), ( Kang Wook Lee ), ( Jae Yong Park ), ( Ji Yeon Chun ), ( Jae Ho Cha ), ( Young Sun Song ), ( Jeong Hwan Kim ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.2
Stromal cell-derived factor-1 (SDF-1/CXCL12) enhances the survival of hematopoietic stem cells, progenitor cells, and a human progenitor cell line MO7e, especially in synergy with other cytokines. Previously, we reported that c-Fos was synergistically induced by combined stimulation with SDF-1/CXCL12 plus other cytokines. Here, we aimed to evaluate that induced c-Fos expression is required for the survival enhancement in MO7e cells. First by using RNase protection assay, we confirmed that synergistic induction of c-Fos was unique among several stress-related genes. Since c-Fos is known as one of the components of transcription factor AP-1, we performed electrophoretic mobility shift assay. The DNA binding activity of AP-1 was also synergistically enhanced by combined stimulation, and c-Fos was identified in the DNA bound complex. To further evaluate the significance of c-Fos expression for survival of MO7e cells, we employed cell permeable c-Fos dominant negative mutant form (A-Fos-MTS) and protein transduction procedure. Inhibition of c-Fos by treatment with A-Fos-MTS significantly reduced survival of MO7e cells in the presence of SDF-1/CXCL12 plus low concentration of GM-CSF. Therefore, we suggest that induction of c-Fos and enhanced AP-1 activity contribute to survival effect of cytokines in MO7e cells.
Bacillus amyloliquefaciens CH86-1 isolated from cheonggukjang was found to have strong fibrinolytic activity when grown on Luria-Bertani medium, and this activity increased sharply when the cells entered the stationary phase. The major fibrinolytic enzyme
A variety of microbes grow by respiration with dimethyl sulfoxide (DMSO) as an electron acceptor, and several distinct DMSO respiratory systems, consisting of electron carriers and a terminal DMSO reductase, have been characterized. The heterotrophic growth of a hyperthermophilic archaeon Thermococcus onnurineus NA1 was enhanced by the addition of DMSO, but the archaeon was not capable of reducing DMSO to DMS directly using a DMSO reductase. Instead, the archaeon reduced DMSO via a cysteine-cystine redox shuttle through a mechanism whereby cystine is microbially reduced to cysteine, which is then reoxidized by DMSO reduction. A thioredoxin reductase-protein disulfide oxidoreductase redox couple was identified to have intracellular cystine-reducing activity, permitting recycle of cysteine. This study presents the first example of DMSO reduction via an electron shuttle. Several Thermococcales species also exhibited enhanced growth coupled with DMSO reduction, probably by disposing of excess reducing power rather than conserving energy.
E-cadherin and its associated cytoplasmic proteins, β-catenin play an essential role in the maintenance of intercellular adhesion in epithelial tissues. It is known that loss or abnormal localization of E-cadherin/catenin from the cell membrane to the cytoplasm correlates with a less favourable clinical outcome and malignancies to human. Thus we evaluated the expression pattern of E-cadherin and β-catenin in gastric carcinoma and analyzed their relationship with tumour clinicopathologic factors and patient survival. Immunohistochemical staining of E-cadherin and β-catenin was performed on 111 paraffin-embedded specimens of gastric carcinoma using an indirect immunoperoxidase technique. Normal membrane staining was observed in 69.4% and 68% gastric carcinoma for E-cadherin and β-catenin, respectively. Abnormal expression of E-cadherin and β-catenin was demonstrated in 30.6% and 38.7% of gastric carcinoma, respectively. Univariate analysis using the Kaplan-Meier showed that the prognostic factors were TNM stage and LN (p<0.05). but E-cadherin and β-catenin was not significant. There was a correlation between E-cadherin expression and β-catenin expression. Multivariate analyses by Cox regression model showed that the independant prognostic factors influencing survival was only TNM stage. Further studies are necessary to assess the role of E-cadherin and β-catenin for the identification of individuals with an increased risk of developing gastric cancer and their clinical management.
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Acne is one of the most common dermatological conditions, but the detailed pathology is unclear and current treatment regimens are plagued by side effects. Cutibacterium acnes is known as a major acne associated bacterium that derives energy from lipase mediated sebum lipid degradation. C. acnes is commensal, but free fatty acids digested by C. acnes lipases from triglyceride and lysophosphatidyl choline induce marked inflammation. Up to date, it remains to be elucidated the lipase activity mechanism at the molecular level. In this study, we report the purification and crystallization of the fulllength lysophospholipase from C. acnes. The crystal diffracted X-rays to a 1.7 Å resolution, revealing that the crystals belong to space group P21212, with cell dimension parameters of a = 94.5, b = 129.7, and c = 55.5 Å. Two protomers are present in the crystallographic asymmetric unit, with a calculated crystal volume per protein weight (VM) of 2.50 Å3Da-1 and a solvent content of 50.79%. This structure will contribute towards revealing a detailed mechanism of acne development.
Zearalenone (ZEA) is an estrogenic mycotoxin that is produced by several Fusarium species, including Fusarium graminearum. One of the ZEA biosynthetic genes, ZEB2, encodes two isoforms of Zeb2 by alternative transcription, forming an activator (Zeb2L-Zeb2L homooligomer) and an inhibitor (Zeb2L-Zeb2S heterodimer) that directly regulate the ZEA biosynthetic genes in F. graminearum. Cyclic AMP-dependent protein kinase A (PKA) signaling regulates secondary metabolic processes in several filamentous fungi. In this study, we investigated the effects of the PKA signaling pathway on ZEA biosynthesis. Through functional analyses of PKA catalytic and regulatory subunits (CPKs and PKR), we found that the PKA pathway negatively regulates ZEA production. Genetic and biochemical evidence further demonstrated that the PKA pathway specifically represses ZEB2L transcription and also takes part in posttranscriptional regulation of ZEB2L during ZEA production. Our findings reveal the intriguing mechanism that the PKA pathway regulates secondary metabolite production by reprograming alternative transcription.