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[en] hnRNP K is a highly conserved nucleocytoplasmic shuttling protein, which associates with RNAs through synergistic binding via its three KH domains. hnRNP K is required for proper nuclear export and translational control of its mRNA targets, and these processes are controlled by hnRNP K's movement between subcellular compartments. Whereas the nuclear export and localization of hnRNP K that is associated with mRNP complexes has been well studied, the trafficking of hnRNP K that is unbound to mRNA has yet to be elucidated. To that end, we expressed an EGFP-tagged RNA binding-defective form of hnRNP K in intact Xenopus embryos, and found it was rapidly degraded in vivo. Deleting hnRNP K's nuclear localization signal (NLS), which contains two prospective ubiquitination sites, rescued the protein from degradation. These data demonstrate a novel activity for the NLS of hnRNP K in regulating the protein's stability in vivo when it is unbound to nucleic acids. - Highlights: • An RNA-binding mutant of Xenopus hnRNP K is degraded in vivo. • hnRNP K's nuclear localization signal contains two predicted ubiquitination sites. • Deletion of the nuclear localization signal rescues RNA-binding mutant degradation.
[en] Activation-induced cytidine deaminase (AID) is essential for diversification of the Ig variable region (IgV). AID is excluded from the nucleus, where it normally functions. However, the molecular mechanisms responsible for regulating AID localization remain to be elucidated. The SR-protein splicing factor SRSF1 is a nucleocytoplasmic shuttling protein, a splicing isoform of which called SRSF1-3, has previously been shown to contribute to IgV diversification in chicken DT40 cells. In this study, we examined whether SRSF1-3 functions in IgV diversification by promoting nuclear localization of AID. AID expressed alone was localized predominantly in the cytoplasm. In contrast, co-expression of AID with SRSF1-3 led to the nuclear accumulation of both AID and SRSF1-3 and the formation of a protein complex that contained them both, although SRSF1-3 was dispensable for nuclear import of AID. Expression of either SRSF1-3 or a C-terminally-truncated AID mutant increased IgV diversification in DT40 cells. However, overexpression of exogenous SRSF1-3 was unable to further enhance IgV diversification in DT40 cells expressing the truncated AID mutant, although SRSF1-3 was able to form a protein complex with the AID mutant. These results suggest that SRSF1-3 promotes nuclear localization of AID probably by forming a nuclear protein complex, which might stabilize nuclear AID and induce IgV diversification in an AID C-terminus-dependent manner. - Highlights: • SRSF1-3 promotes the nuclear accumulation of AID. • SRSF1-3 forms a protein complex with AID in an AID C-terminus-independent manner. • SRSF1-3 contributes to IgV hypermutation in an AID C-terminus-dependent manner.
[en] Hypoxia-dependent gene regulation is largely orchestrated by hypoxia-inducible factors (HIFs), which associate with defined nucleotide sequences of hypoxia-responsive elements (HREs). Comparison of the regulatory HRE within the 3′ enhancer of the human erythropoietin (EPO) gene with known binding motifs for cold shock protein Y-box (YB) protein-1 yielded strong similarities within the Y-box element and 3′ adjacent sequences. DNA binding assays confirmed YB-1 binding to both, single- and double-stranded HRE templates. Under hypoxia, we observed nuclear shuttling of YB-1 and co-immunoprecipitation assays demonstrated that YB-1 and HIF-1α physically interact with each other. Cellular YB-1 depletion using siRNA significantly induced hypoxia-dependent EPO production at both, promoter and mRNA level. Vice versa, overexpressed YB-1 significantly reduced EPO-HRE-dependent gene transcription, whereas this effect was minor under normoxia. HIF-1α overexpression induced hypoxia-dependent gene transcription through the same element and accordingly, co-expression with YB-1 reduced HIF-1α-mediated EPO induction under hypoxic conditions. Taken together, we identified YB-1 as a novel binding factor for HREs that participates in fine-tuning of the hypoxia transcriptome. - Highlights: • Hypoxia drives nuclear translocation of cold shock protein YB-1. • YB-1 physically interacts with hypoxia-inducible factor (HIF)-1α. • YB-1 binds to the hypoxia-responsive element (HRE) within the erythropoietin (EPO) 3′ enhancer. • YB-1 trans-regulates transcription of hypoxia-dependent genes such as EPO and VEGF.
[en] Base on the planning to increase of the working capability of the working capability of the rabbit system facility, the design of mentioned portable bridge on access has been done. Using the designed installation the transportation of the middle and high active samples from the isotop cell to the scanning room will be easy to be done. The design has been done by using the maximum actual load. Determination of maximum actual load, determination of maximum actual load, and also the chosen steel shape dimension compliance with ASTM standard. The installation required 2 peaces of W 6 x 12 steel beam by 111.82 inches in length, 4 peaces W 6 x 12 steel beam by 47.25in length and a peace of steel plate by 111.82x 47.25x 0,394. This paper concluded that this design is feasible to be fabricated
[en] The rabbit transfer system facility is a facility that a function to decrease the transfer times of the irradiated samples from isotop cell to counting room. Base on the planning to increase the research activity and the service quality to the rabbit system user's is necessary to realized manufacturing and installation of the mention facility. The detail design purpose needs 35 m length of polypinil hose by 36 mm inner diameter and 42 mm outer diameter. By doing analysis the transfer time needs to move the sample is 3 seconds. As a reference use ASME, ASTM and AISC standard
[en] Micron-sized objects having asymmetric boundaries can rectify the chaotic motions of an active bacterial suspension and perform geometrically biased random walks. Using numerical simulations in a planar geometry, we show that arrow-shaped micro-shuttles, constrained to move in one dimension (1D) in a bath of self-propelled micro-organisms, spontaneously perform unidirectional translational motions with a strongly shape-dependent speed. Relaxing the 1D constraint, a random motion in the whole plane sets in at long times, due to random changes in shuttle orientation caused by bacterial collisions. The complex dynamics arising from the mechanical interactions between bacteria and the object boundaries can be described by a Gaussian stochastic force with a shape-dependent mean and a self-correlation decaying exponentially on the timescale of seconds.
[en] A quantum shuttle is an archetypical nanoelectromechanical device, where the mechanical degree of freedom is quantized. Using a full-scale numerical solution of the generalized Master equation describing the shuttle, we have recently shown (Novotny et al 2004 Phys. Rev. Lett. 92 248302) that for certain limits of the shuttle parameters one can distinguish three distinct charge transport mechanisms: (i) an incoherent tunnelling regime (ii) a shuttling regime, where the charge transport is synchronous with the mechanical motion, and (iii) a coexistence regime, where the device switches between the tunnelling and shuttling regimes. While a study of the crossover between these three regimes requires the full numerics, we show here that by identifying the appropriate timescales it is possible to derive vastly simpler equations for each of the three regimes. The simplified equations allow a clear physical interpretation, are easily solved and are in good agreement with the full numerics in their respective domains of validity
[en] This paper presents the design, fabrication and calibration of a novel MEMS logic gate that can perform Boolean algebra as well as logic devices composed of solid-state transistors. Unlike existing designs, the proposed design can perform either NAND gate or NOR gate functions using the same mechanical structure, but different electrical interconnects. The proposed design imposes three requirements on the fabrication process: two voltage levels carried on a suspended plate, metal-to-metal contact between shuttle electrodes and fixed electrodes, and a low process temperature (<300 °C). To fulfill these requirements, the residual stress in the fabricated device is substantial which could impair the functionality of the device. Therefore, a novel in situ film stress calibration method is developed to assist the development of the fabrication process. In a prototype design, the fabricated device is 250 μm long, 100 μm wide and of 3.97 μm gap. Experimental results show that the device can operate at 25/−25 V and 100 Hz, and achieve the proposed logic functions. In addition, several properties of this device are experimentally evaluated, including power consumption, on/off resistance, lifetime and resonant frequency.
[en] The dynamical localization phenomena in two-electron quantum-dot shuttles driven by an ac field have been investigated and analyzed by the Floquet theory. The dynamical localization occurs near the anti-crossings in Floquet eigenenergy spectrum. The oscillation of the quantum-dot shuttles may increase the possibility of the dynamical localization. Especially, even if the two electrons are initialized in two neighbor dots, they can be localized there for appropriate intensity of the driven field. The studies may help the understanding of dynamical localization in electron shuttles and expand the application potential of nanoelectromechanical devices. -- Highlights: ► The dynamical localization in electron shuttle is studied by Floquet theory. ► There is a relation between quasi-energy anti-crossings and dynamical localization. ► The oscillation of quantum dot increases the dynamical localization. ► Even the electrons are initialized in different dots, the localization can occur.