[en] Highlights: • Cav-1 knockdown promoted the expression of miR-145, miR-124 and miR-183. • Cav-1 knockdown inhibited the nuclear translocation of EGR1. • Cav-1 knockdown reduced the interaction between EGR1 and KLF5. • Free KLF5 positively regulated the expression of miR-145, miR-124 and miR-183. • miR-145, miR-124 and miR-183 reduced pituitary adenoma cells migration and invasion. Caveolin-1 (Cav-1) is a principal structural protein of caveolae. Cav-1 has been implicated in cancer progression, but its precise functional roles in pituitary adenoma cells remain largely unclear. In this study, we evidenced that the level of cav-1 was elevated in the invasive pituitary adenoma. Cav-1 knockdown restrained the migration and invasion of pituitary adenoma cells. In cav-1-depleting cells, the expression of miR-145, miR-124 and miR-183 were up-regulated. Further investigation showed that cav-1 knockdown inhibited the nuclear translocation of EGR1, reducing the interaction between EGR1 and KLF5. The resulting free KLF5 promoted the expression of miR-145, miR-124 and miR-183 by binding to their promoters, which was blocked by EGR1. Luciferase reporter assay indicated that miR-145 targeted FSCN1, miR-124 targeted PTTG1IP, and miR-183 targeted EZR in pituitary adenoma cells, respectively. Knockdown of FSCN1, PTTG1IP or EZR suppressed the migration and invasion of pituitary adenoma cells. In conclusion, our data suggested that the elevated cav-1 promoted pituitary adenoma cells migration and invasion by regulating the interaction between EGR1 and KLF5.

[en] Paramyxoviruses have been shown to produce proteins that inhibit interferon production and signaling. For human respiratory syncytial virus (RSV), the nonstructural NS1 and NS2 proteins have been shown to have interferon antagonist activity through an unknown mechanism. To understand further the functions of NS1 and NS2, we generated recombinant RSV in which both NS1 and NS2 were replaced by the PIV5 V protein, which has well-characterized IFN antagonist activities (ΔNS1/2-V). Expression of V was able to partially inhibit IFN responses in ΔNS1/2-V-infected cells. In addition, the replication kinetics of ΔNS1/2-V were intermediate between ΔNS1/2 and wild-type (rA2) in A549 cells. However, expression of V did not affect the ability of ΔNS1/2-V to activate IRF3 nuclear translocation and IFNβ transcription. These data indicate that V was able to replace some of the IFN inhibitory functions of the RSV NS1 and NS2 proteins, but also that NS1 and NS2 have functions in viral replication beyond IFN antagonism

No abstract available

[en] In this study, we investigated the relative participation of N-ethylmaleimide-sensitive factor (NSF) in vivo in a complex multistep vesicle trafficking system, the translocation response of GLUT4 to insulin in rat adipose cells. Transfections of rat adipose cells demonstrate that over-expression of wild-type NSF has no effect on total, or basal and insulin-stimulated cell-surface expression of HA-tagged GLUT4. In contrast, a dominant-negative NSF (NSF-D1EQ) can be expressed at a low enough level that it has little effect on total HA-GLUT4, but does reduce both basal and insulin-stimulated cell-surface HA-GLUT4 by ∼50% without affecting the GLUT4 fold-translocation response to insulin. However, high expression levels of NSF-D1EQ decrease total HA-GLUT4. The inhibitory effect of NSF-D1EQ on cell-surface HA-GLUT4 is reversed when endocytosis is inhibited by co-expression of a dominant-negative dynamin (dynamin-K44A). Moreover, NSF-D1EQ does not affect cell-surface levels of constitutively recycling GLUT1 and TfR, suggesting a predominant effect of low-level NSF-D1EQ on the trafficking of GLUT4 from the endocytic recycling compared to the intracellular GLUT4-specific compartment. Thus, our data demonstrate that the multiple fusion steps in GLUT4 trafficking have differential quantitative requirements for NSF activity. This indicates that the rates of plasma and intracellular membrane fusion reactions vary, leading to differential needs for the turnover of the SNARE proteins

[en] Research highlights: → Vimentin is shown to bind to the N-terminus of insulin-responsive aminopeptidase (IRAP), a major cargo protein of GLUT4 vesicles in 3T3-L1 adipocytes. → GLUT4 translocation to the plasma membrane by insulin is decreased in vimentin-depleted adipocytes. → An interaction between vimentin and IRAP functions to sequester GLUT4 vesicles to the peri-nuclear region of the cell. -- Abstract: Insulin-responsive aminopeptidase (IRAP) and GLUT4 are two major cargo proteins of GLUT4 storage vesicles (GSVs) that are translocated from a postendosomal storage compartment to the plasma membrane (PM) in response to insulin. The cytoplasmic region of IRAP is reportedly involved in retention of GSVs. In this study, vimentin was identified using the cytoplasmic domain of IRAP as bait. The validity of this interaction was confirmed by pull-down assays and immunoprecipitation in 3T3-L1 adipocytes. In addition, it was shown that GLUT4 translocation to the PM by insulin was decreased in vimentin-depleted adipocytes, presumably due to dispersing GSVs away from the cytoskeleton. These findings suggest that the IRAP binding protein, vimentin, plays an important role in retention of GSVs.

[en] In this study, TNF-α was found to activate the WNT/β-catenin pathway in BEAS-2B human bronchial epithelial cells. Levels of phospho-LRP6, Dvl-2, and phospho-GSK-3β were elevated, while that of Axin was reduced by TNF-α treatment. Nuclear translocation of β-catenin and the reporter activity of a β-catenin-responsive promoter were increased by TNF-α treatment. Under the same experimental conditions, TNF-α activated the NF-κB signaling, which includes the phosphorylation and degradation of IκB and nuclear translocation and target DNA binding of NF-κB, and it was found that an inhibitor of NF-κB activation, JSH-23, inhibited TNF-α-induced Wnt signaling as well as NF-κB signaling. It was also found that recombinant Wnt proteins induced NF-κB nuclear translocations and its target DNA binding, suggesting that Wnt signaling and NF-κB signaling were inter-connected. TNF-α-induced modulations of IκB and NF-κB as well as pro-inflammatory cytokine expression were significantly suppressed by the transfection of β-catenin siRNA compared to that of control siRNA. Transfection of a β-catenin expression plasmid augmented the TNF-α-induced modulations of IκB and NF-κB as well as pro-inflammatory cytokine expression. These results clearly demonstrated that the WNT/β-catenin pathway modulates the inflammatory response induced by TNF-α, suggesting that this pathway may be a useful target for the effective treatment of bronchial inflammation. - Highlights: • TNF-α activated the WNT/β-catenin pathway in bronchial epithelial cells. • TNF-α-induced WNT/β-catenin pathway was inhibited by NF-κB inhibitor. • Wnt proteins induced nuclear translocation and target DNA binding of NF-κB. • TNF-α-induced NF-κB signaling was suppressed by β-catenin knock-down. • TNF-α-induced NF-κB signaling was augmented by ectopic β-catenin expression.

[en] We study pore-blockade times for a translocating polymer of length N, driven by a field E across the pore in three dimensions. The polymer performs Rouse dynamics, i.e., we consider polymer dynamics in the absence of hydrodynamical interactions. We find that the typical time for which the pore remains blocked during a translocation event scales as ∼N^{(1+2ν)/(1+ν)}/E, where ν approx. = 0.588 is the Flory exponent for the polymer. We show, in line with our previous work, that this scaling behavior stems from polymer dynamics in the immediate vicinity of the pore-in particular, the memory effects in the polymer chain tension imbalance across the pore. This result, like numerical results from several other groups, violates the lower bound ∼N^1+ν/E suggested earlier in the literature. We discuss why this lower bound is incorrect and show, on the basis of the conservation of energy, that the correct lower bound for the pore-blockade time for field-driven translocation is given by ηN^2ν/E, where η is the viscosity of the medium surrounding the polymer

[en] The paper presents results of the regional background frequency of stable translocations using in situ fluorescent hybridization in different age groups, which ranges from 1.42±0.3 to 4.9±0.5 per 1000 cells. It found that frequency of stable chromosome aberrations revealed by FISH-technique increases with age. This is due to the fact that with age chromosome stability, DNA reparation and regeneration reduction progressively decelerate. The impact of bad habits such as smoking is the most studied sign affecting the level of translocations; therefore, correlation was made between the level of stable chromosome damage and this bad habit in age groups of interest. Background frequency of stable chromosome damage was calculated by means of equipment of automated cytogenetic platform based on the electronic Carl Zeiss AxioImager Z2 fluorescent microscope, automatic Metafer 4/M Search, ISIS (MetaSystems, Germany) metaphase search and analysis system and using commercial whole chromosome DNA probes for chromosomes 1, 4, 12.

[en] The capture and translocation of biomolecules through nanometer-scale pores are processes with a potentially large number of applications, and hence they have been intensively studied in recent years. The aim of this paper is to review existing models of the capture process by a nanopore, together with some recent experimental data of short single- and double-stranded DNA captured by the Cytolysin A (ClyA) nanopore. ClyA is a transmembrane protein of bacterial origin which has been recently engineered through site-specific mutations, to allow the translocation of double- and single-stranded DNA. A comparison between theoretical estimations and experiments suggests that for both cases the capture is a reaction-limited process. This is corroborated by the observed salt dependence of the capture rate, which we find to be in quantitative agreement with the theoretical predictions. (paper)

[en] IL-33 is a member of the IL-1 family of cytokines with dual function which either activates cells via the IL-33 receptor in a paracrine fashion or translocates to the nucleus to regulate gene transcription in an intracrine manner. We show that full length murine IL-33 is active as a cytokine and that it is not processed by caspase 1 to mature IL-33 but instead cleaved by caspase 3 at aa175 to yield two products which are both unable to bind to the IL-33 receptor. Full length IL-33 and its N-terminal caspase 3 breakdown product, however, translocate to the nucleus. Finally, bioactive IL-33 is not released by cells constitutively or after activation. This suggests that IL-33 is not a classical cytokine but exerts its function in the nucleus of intact cells and only activates others cells via its receptor as an alarm mediator after destruction of the producing cell.