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The Stable Association of Virion with the Triple-geneblockProtein 3-based Complex of Bamboo mosaic virus
The triple-gene-block protein 3 (TGBp3) of Bamboo mosaic virus (BaMV) is an integral endoplasmic reticulum (ER) membraneprotein which is assumed to form a membrane complex to deliver the virus intracellularly. However, the virus entity that isdelivered to plasmodesmata (PD) and its association with TGBp3-based complexes are not known. Results from chemicalextraction and partial proteolysis of TGBp3 in membrane vesicles revealed that TGBp3 has a right-side-out membranetopology; i.e., TGBp3 has its C-terminal tail exposed to the outer surface of ER. Analyses of the TGBp3-specificimmunoprecipitate of Sarkosyl-extracted TGBp3-based complex revealed that TGBp1, TGBp2, TGBp3, capsid protein (CP),replicase and viral RNA are potential constituents of virus movement complex. Substantial co-fractionation of TGBp2, TGBp3and CP, but not TGBp1, in the early eluted gel filtration fractions in which virions were detected after TGBp3-specificimmunoprecipitation suggested that the TGBp2- and TGBp3-based complex is able to stably associate with the virion. Thisnotion was confirmed by immunogold-labeling transmission electron microscopy (TEM) of the purified virions. In addition,mutational and confocal microscopy analyses revealed that TGBp3 plays a key role in virus cell-to-cell movement byenhancing the TGBp2- and TGBp3-dependent PD localization of TGBp1. Taken together, our results suggested that the cellto-cell movement of potexvirus requires stable association of the virion cargo with the TGBp2- and TGBp3-based membranecomplex and recruitment of TGBp1 to the PD by this complex
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Pluripotency-associated genes in human nasopharyngeal carcinoma CNE-2 cells are reactivated by a unique epigenetic sub-microenvironment.
RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.BACKGROUND: There is increasing evidence that cancers contain their own stem-like cells, and particular attention has been paid to one subset of cancer-stem cells termed side population (SP). Stem cells under normal physical conditions are tightly controlled by their microenvironment, however, the regulatory role of the microenvironment surrounding cancer stem cells is not well characterized yet. In this study we found that the phenotype of SP can be "generated" by macrophage-like cells under conditioned culture. Furthermore the gene regulation pathway involved in cellular reprogramming process was investigated. METHODS: The selection and identification of SP in 50 CNE-2 single cell clones were performed by flow cytometry. The transwell assay and immunofluorescence staining were used to measure migration and cancer stem cell characters of non-SP single clone cells cultured with conditioned medium respectively. The subtraction suppression hybridization (SSH) technique and northern blotting analysis was applied to explore the pluripotency-associated genes under a unique epigenetic sub-microenvironment. RESULTS: Among 50 clones, only one did not possess SP subpopulation while others did. The non-SP cells induced by macrophage-like cells showed more aggressive characters, which increased cell migration compared with the control cells and showed some fraction of SP phenotype. These cells expressed distinguished level of pluripotency-associated genes such as ADP-ribosylation factor-like 6 interacting protein (ARMER), poly (rC) binding protein 1 (PCBP1) and pyruvate dehydrogenase E1-beta subunit (PDHB) when subjected to the environment. CONCLUSION: To our knowledge, this is the first study to demonstrate that non-SP single-clone cells can be induced to generate a SP phenotype when they are cultured with conditioned medium of macrophage-like cells, which is associated with the reactivation of pluripotency-associated genes.Peer Reviewe
Comparative analysis of the secretomes of Schizophyllum commune and other wood-decay basidiomycetes during solid-state fermentation reveals its unique lignocellulose-degrading enzyme system
Additional file 3: Table S2. Identified proteins in the secretomes of four fungi during SSF on Jerusalem artichoke stalk
Flood Damage Assessment in Taipei City Taiwan
In this study, we reviewed the literature on flood damage assessment and collected information for related research in Taiwan to analyze the relationships between direct flood damage, flood frequency, flood depth, and land-use. The procedure for flood damage assessment was then developed that includes the following steps: (a) Scenario simulation of inundation potential. (b) Establishment of the relationship between inundation depth and damage loss for varied land-use. (c) Risk analysis of inundation damage.
Taipei City in north Taiwan was adopted as the case study to demonstrate the proposed algorithm. Flood events with return periods of 5, 10, 25, 50, 100 and 200 years were used for flood hazard analysis to cover possible flooding scenarios. The inundation hazard maps were first generated via hydraulic modelling. The regional flood damage was then estimated using a relationship between inundation depth and damage. The flood damage exceedance probability (EP) curve for Taipei City was constructed following the association of the loss with its probability of occurrence. The flood damage EP curve was further used to integrate the damage assessments for individual flood events for a full probability range presentation of the flood risk. The expected annual damage was calculated by integrating the area under the EP curve
SEC24A deficiency lowers plasma cholesterol through reduced PCSK9 secretion.
The secretory pathway of eukaryotic cells packages cargo proteins into COPII-coated vesicles for transport from the endoplasmic reticulum (ER) to the Golgi. We now report that complete genetic deficiency for the COPII component SEC24A is compatible with normal survival and development in the mouse, despite the fundamental role of SEC24 in COPII vesicle formation and cargo recruitment. However, these animals exhibit markedly reduced plasma cholesterol, with mutations in Apoe and Ldlr epistatic to Sec24a, suggesting a receptor-mediated lipoprotein clearance mechanism. Consistent with these data, hepatic LDLR levels are up-regulated in SEC24A-deficient cells as a consequence of specific dependence of PCSK9, a negative regulator of LDLR, on SEC24A for efficient exit from the ER. Our findings also identify partial overlap in cargo selectivity between SEC24A and SEC24B, suggesting a previously unappreciated heterogeneity in the recruitment of secretory proteins to the COPII vesicles that extends to soluble as well as trans-membrane cargoes. DOI:http://dx.doi.org/10.7554/eLife.00444.001
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