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At least 1,603 records · Page 89

Regulation of RNA-binding proteins affinity to export receptors enables the nuclear basket proteins to distinguish and retain aberrant mRNAs

Export of messenger ribonucleic acids (mRNAs) into the cytoplasm is a fundamental step in gene regulation processes, which is meticulously quality controlled by highly efficient mechanisms in eukaryotic cells. Yet, it remains unclear how the aberrant mRNAs are recognized and retained inside the nucleus. Using a new modelling approach for complex systems, namely the agent-based modelling (ABM) approach, we develop a minimal model of the mRNA quality control (QC) mechanism. Our results demonstrate that regulation of the affinity of RNA-binding proteins (RBPs) to export receptors along with the weak interaction between the nuclear basket protein (Mlp1 or Tpr) and RBPs are the minimum requirements to distinguish and retain aberrant mRNAs. Our results show that the affinity between Tpr and RBPs is optimized to maximize the retention of aberrant mRNAs. In addition, we demonstrate how the length of mRNA affects the QC process. Since longer mRNAs spend more time in the nuclear basket to form a compact conformation and initiate their export, nuclear basket proteins could more easily capture and retain them inside the nucleus.

Science & Technology - Other Topics↗

Regulating electrodeposition morphology of lithium: towards commercially relevant secondary Li metal batteries

Lithium, the lightest and most electronegative metallic element, has long been considered the ultimate choice as a battery anode for mobile, as well as in some stationary applications. The high electronegativity of Li is, however, a double-edged sword—it facilitates a large operating voltage when paired with essentially any cathode, promising a high cell-level energy density. It is also synonymous with a high chemical reactivity and low reduction potential. The interfaces a Li metal anode forms with any other material (liquid or solid) in an electrochemical cell are therefore always mediated by one or more products of its chemical or electrochemical reactions with that material. The physical, crystallographic, mechanical, electrochemical, and transport properties of the resultant new material phases (interphases) regulate all interfacial processes at a Li metal anode, including electrodeposition during battery recharge. This Review takes recent efforts aimed at manipulating the structure, composition, and physical properties of the solid electrolyte interphase (SEI) formed on an Li anode as a point of departure to discuss the structural, electrokinetic, and electrochemical requirements for achieving high anode reversibility. An important conclusion is that while recent reports showing significant advances in the achievement of highly reversible Li anodes, e.g. as measured by the coulombic efficiency (CE), raise prospects for as significant progress towards commercially relevant Li metal batteries, the plateauing of achievable CE values to around 99 ± 0.5% apparent from a comprehensive analysis of the literature is problematic because CE values of at least 99.7%, and preferably >99.9% are required for Li metal cells to live up to the potential for higher energy density batteries offered by the Li metal anode. On this basis, we discuss promising approaches for creating purpose-built interphases on Li, as well as for fabricating advanced Li electrode architectures for regulating Li electrodeposition morphology and crystallinity. Considering the large number of physical and chemical factors involved in achieving fine control of Li electrodeposition, we believe that achievement of the remaining ~0.5% in anode reversibility will require fresh approaches, perhaps borrowed from other fields. We offer perspectives on both current and new strategies for achieving such Li anodes with the specific aim of engaging established contributors and newcomers to the field in the search for scalable solutions.

25 ENERGY STORAGE↗

Electrochemical pH regulation in droplet microfluidics

Here we report a method for electrochemical pH regulation in microdroplets generated in a microfluidic device. The key finding is that controlled quantities of reagents can be generated electrochemically in moving microdroplets confined within a microfluidic channel. Additionally, products generated at the anode and cathode can be isolated within descendant microdroplets. Specifically, ~5 nL water-in-oil microdroplets are produced at a T-junction and then later split into two descendant droplets. During splitting, floor-patterned microelectrodes drive water electrolysis within the aqueous microdroplets to produce H + and OH - . This results in a change in the pHs of the descendant droplets. The droplet pH can be regulated over a range of 5.9 to 7.7 by injecting controlled amounts of charge into the droplets. When the injected charge is between -6.3 and 54.5 nC nL -1 , the measured pH of the resulting droplets is within ±0.1 pH units of that predicted based on the magnitude of the injected charge. This technique can likely be adapted to electrogeneration of other reagents within microdroplets.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Brush-modified fluorescent organic nanoparticles by ATRP with rigidity-regulated emission

Organic nanoparticles provide exceptional intraparticle tailorability, enabling the incorporation of functional molecules for diverse applications. In this study, we present the synthesis and characterization of fluorescent organic nanoparticles (FoNPs) with encapsulated aggregation-induced emission (AIE) luminogens with emission properties regulated by particle rigidity. Atom transfer radical polymerization (ATRP) was employed in dispersed media to develop various fluorescence colors tuned by precise control over particle rigidity. Comprehensive analyses revealed that increased particle rigidity significantly enhanced photoluminescence, achieving quantum yields of up to 22% in selected solvents. The high chain-end fidelity facilitated the grafting of hydrophilic polymer brushes from surfaces of FoNPs used as macroinitiators, enabling their dispersion in aqueous media while maintaining bright fluorescence. These findings highlight the potential of rigidity-regulated FoNPs as versatile platforms for advanced material applications, particularly in fluorescent waterborne films and aqueous-phase sensing systems.

Yin, Rongguan [Carnegie Mellon Univ., Pittsburgh, ↗

Coordinative Real-time Sub-Transmission Volt-VAr Control for Reactive Power Regulation between Transmission and Distribution Systems

This paper introduces an integrated voltage regulation approach to coordinate the Volt-VAr control between transmission and distribution systems. Distributed photovoltaic (PV) systems are aggregated as a virtual power plant (VPP). At the sub-transmission level, a Coordinated Real-time Sub-Transmission Volt-VAr Control Tool (CReST-VCT) is introduced to coordinate the operation of the shunt devices and the reactive power provided by the VPPs. The CReST-VCT uses an enhanced Volt-VAr control algorithm with weighted voltage control objects that include minimizing losses, maintaining the voltage within desirable levels, and minimizing the number of mechanical switching of shunt elements. The algorithm runs every 5 minutes and is solved using AC optimal power flow (OPF) solution technique. The outputs include the settings for each sub-transmission-level Volt-VAr control devices and the reactive power generation requirements for each VPP. Then, each VPP will run a distribution Volt-VAr control algorithm to select the PV inverters that provide the Volt-VAr control services considering the distribution network topology. The goal of the VPP reactive power control is to meet the transmission level service requirements while maintaining the voltage and line flow in the distribution system within limits. Each VPP will update its voltage regulation capacity every 5 minutes to allow the CReST-VCT to formulate the optimization problem for the next dispatch interval. The proposed tool is simulated on Duke Energy Carolina system to demonstrate the capability of providing voltage support service by dispatching of reactive power of distributed PV as VPP.

Ke, Xinda↗

Arc discharge regulation of a megawatt hot cathode bucket ion source for the experimental advanced superconducting tokamak neutral beam injector

Arc discharge of a hot cathode bucket ion source tends to be unstable what attributes to the filament self-heating and energetic electrons backstreaming from the accelerator. A regulation method, which based on the ion density measurement by a Langmuir probe, is employed for stable arc discharge operation and long pulse ion beam generation. Long pulse arc discharge of 100 s is obtained based on this regulation method of arc power. It establishes a foundation for the long pulse arc discharge of a megawatt ion source, which will be utilized a high power neutral beam injection device.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Charge regulation and local dielectric function in planar polyelectrolyte brushes

Understanding the effect of inhomogeneity on the charge regulation and dielectric properties, andhow it depends on the conformational characteristics of the macromolecules is a long-standing problem. In order to address this problem, we have developed a field-theory to study charge regulation and local dielectric function in planar polyelectrolyte brushes. The theory is used to study a polyacid brush, which is comprised of chains end-grafted at the solid-fluid interface, in equilibrium with a bulk solution containing monovalent salt ions, solvent molecules, and pH controlling acid. In particular, we focus on the effects of the concentration of added salt and pH of the bulk in determining the local charge and dielectric function. Our theoretical investigations reveal that the dipole moment of the ion-pairs formed as a result of counterion adsorption on the chain backbones play a key rolein affecting the local dielectric function. For polyelectrolytes made of monomers having dipole moments lower than the solvent molecules, dielectric decrement is predicted inside the brush region.However, the formation of ion-pairs (due to adsorption of counterions coming from the dissociationof added salt) more polar than the solvent molecules is shown to increase the magnitude of the dielectric function with respect to its bulk value. Furthermore, an increase in the bulk salt concentration is shown to increase the local charge inside the brush region.

36 MATERIALS SCIENCE↗

Note: A new regulation method of stable operation of high power cathode ion source

The hot cathode ion source will tend to be unstable when operated with high power and long pulse. In order to achieve stable operation, a new regulation method based on the arc power (discharge power) feedback control was designed and tested on the hot cathode ion source test bed with arc discharge and beam extraction. The results show that the new regulation method can achieve stable arc discharge and beam extraction. It verifies the success of feedback control of arc source with arc power.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

A miniaturized piezoelectric turbine with self-regulation for increased air speed range

This paper presents the design and demonstration of a piezoelectric turbine with self-regulation for increased air speed range. The turbine's transduction is achieved by magnetic “plucking” of a piezoelectric beam by the passing rotor. The increased speed range is achieved by the self-regulating mechanism which can dynamically adjust the magnetic coupling between the magnets on the turbine rotor and the piezoelectric beam using a micro-spring. The spring is controlled passively by the centrifugal force of the magnet on the rotor. This mechanism automatically changes the relative position of the magnets at different rotational speeds, making the coupling weak at low airflow speeds and strong at high speeds. Hence, the device can start up with a low airflow speed, and the output power can be ensured when the airflow speed is high. A theoretical model was established to analyse the turbine's performance, advantages, and to optimize its design parameters. A prototype was fabricated and tested in a wind tunnel. The start-up airflow speed was 2.34 m/s, showing a 30% improvement against a harvester without the mechanism.

42 ENGINEERING↗

Modeling cutinase enzyme regulation in polyethylene terepthalate plastic biodegradation

PET (Polyethylene terephthalate) is a plastic material that is commonly used in our daily life. The high production of PET and others plastics that can be up to three hundred million tons per year, is not matched by its degradation rate and hence leads to environmental pollution. To overcome this problem, we develop a biodegradation system. This system utilizes LC Cutinase enzyme produced by engineered escherichia coli bacteria to degrade PET. To make the system works efficaciously, it is important to understand the mechanism underlying its enzyme regulation. Therefore, we construct a mathematical model to describe the regulation of LC Cutinase production. The stability of the model is analyzed. We show that the designated biodegradation system can give an oscillatory behavior that is very important to control the amount of inclusion body (the miss-folded proteins that reduce the efficiency of the biodegradation system).

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Self-regulated growth of candidate topological superconducting parkerite by molecular beam epitaxy

Ternary chalcogenides, such as parkerites and shandites, are a broad class of materials exhibiting a rich diversity of transport and magnetic behavior and an array of topological phases, including Weyl and Dirac nodes. However, they remain largely unexplored as high-quality epitaxial thin films. Here, we report the self-regulated growth of thin films of the strong spin–orbit coupled superconductor Pd 3 Bi 2 Se 2 on SrTiO 3 by molecular beam epitaxy. Films are found to grow in a self-regulated fashion, where, in excess Se, the temperature and relative flux ratio of Pd to Bi control the formation of Pd 3 Bi 2 Se 2 due to the combined volatility of Bi, Se, and Bi–Se bonded phases. The resulting films are shown to be of high structural quality, and the stoichiometry is independent of the Pd:Bi and Se flux ratio and exhibits a superconducting transition temperature of 800 mK and a critical field of 17.7 ± 0.5 mT, as probed by transport and magnetometry. Understanding and navigating the growth of the chemically and structurally diverse classes of ternary chalcogenides open a vast space for discovering new phenomena and enabling new applications.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Arabidopsis ribosomal proteins control vacuole trafficking and developmental programs through the regulation of lipid metabolism

The vacuole is the most prominent compartment in plant cells and is important for ion and protein storage. In our effort to search for key regulators in the plant vacuole sorting pathway, ribosomal large subunit 4 (rpl4d) was identified as a translational mutant defective in both vacuole trafficking and normal development. Polysome profiling of the rpl4d mutant showed reduction in polysome-bound mRNA compared with wild-type, but no significant change in the general mRNA distribution pattern. Ribsomal profiling data indicated that genes in the lipid metabolism pathways were translationally down-regulated in the rpl4d mutant. Live imaging studies by Nile red staining suggested that both polar and nonpolar lipid accumulation was reduced in meristem tissues of rpl4d mutants. Pharmacological evidence showed that sterol and sphingolipid biosynthetic inhibitors can phenocopy the defects of the rpl4d mutant, including an altered vacuole trafficking pattern. Genetic evidence from lipid biosynthetic mutants indicates that alteration in the metabolism of either sterol or sphingolipid biosynthesis resulted in vacuole trafficking defects, similar to the rpl4d mutant. Tissue-specific complementation with key enzymes from lipid biosynthesis pathways can partially rescue both vacuole trafficking and auxin-related developmental defects in the rpl4d mutant. These results indicate that lipid metabolism modulates auxin-mediated tissue differentiation and endomembrane trafficking pathways downstream of ribosomal protein function.

59 BASIC BIOLOGICAL SCIENCES↗

PLP and GABA trigger GabR-mediated transcription regulation in Bacillus subtilis via external aldimine formation

Here, the Bacillus subtilis protein regulator of the gabTD operon and its own gene (GabR) is a transcriptional activator that regulates transcription of gamma-aminobutyric acid aminotransferase (GABA-AT; GabT) upon interactions with pyridoxal-5'-phosphate (PLP) and GABA, and thereby promotes the biosynthesis of glutamate from GABA. We show here that the external aldimine formed between PLP and GABA is apparently responsible for triggering the GabR-mediated transcription activation. Details of the "active site" in the structure of the GabR effector-binding/oligomerization (Eb/O) domain suggest that binding a monocarboxylic.-amino acid such as GABA should be preferred over dicarboxylic acid ligands. A reactive GABA analog, (S)-4-amino-5-fluoropentanoic acid (AFPA), was used as a molecular probe to examine the reactivity of PLP in both GabR and a homologous aspartate aminotransferase (Asp-AT) from Escherichia coli as a control. A comparison between the structures of the Eb/O-PLP-AFPA complex and Asp-AT-PLP-AFPA complex revealed that GabR is incapable of facilitating further steps of the transamination reaction after the formation of the external aldimine. Results of in vitro and in vivo assays using full-length GabR support the conclusion that AFPA is an agonistic ligand capable of triggering GabR-mediated transcription activation via formation of an external aldimine with PLP.

59 BASIC BIOLOGICAL SCIENCES↗

Structural and functional studies of pyruvate carboxylase regulation by cyclic di-AMP in lactic acid bacteria

Cyclic di-3',5'-adenosine monophosphate (c-di-AMP) is a broadly conserved bacterial second messenger that has been implicated in a wide range of cellular processes. Our earlier studies showed that c-di-AMP regulates central metabolism in Listeria monocytogenes by inhibiting its pyruvate carboxylase (LmPC), a biotin-dependent enzyme with biotin carboxylase (BC) and carboxyltransferase (CT) activities. We report here structural, biochemical, and functional studies on the inhibition of Lactococcus lactis PC (LlPC) by c-di-AMP. The compound is bound at the dimer interface of the CT domain, at a site equivalent to that in LmPC, although it has a distinct binding mode in the LlPC complex. Here, this binding site is not well conserved among PCs, and only a subset of these bacterial enzymes are sensitive to c-di-AMP. Conformational changes in the CT dimer induced by c-di-AMP binding may be the molecular mechanism for its inhibitory activity. Mutations of residues in the binding site can abolish c-di-AMP inhibition. In L. lactis, LlPC is required for efficient milk acidification through its essential role in aspartate biosynthesis. The aspartate pool in L. lactis is negatively regulated by c-di-AMP, and high aspartate levels can be restored by expression of a c-di-AMP–insensitive LlPC. LlPC has high intrinsic catalytic activity and is not sensitive to acetyl-CoA activation, in contrast to other PC enzymes.

59 BASIC BIOLOGICAL SCIENCES↗

An endogenous dAMP ligand in Bacillus subtilis class Ib RNR promotes assembly of a noncanonical dimer for regulation by dATP

The high fidelity of DNA replication and repair is attributable, in part, to the allosteric regulation of ribonucleotide reductases (RNRs) that maintains proper deoxynucleotide pool sizes and ratios in vivo. In class Ia RNRs, ATP (stimulatory) and dATP (inhibitory) regulate activity by binding to the ATP-cone domain at the N terminus of the large α subunit and altering the enzyme’s quaternary structure. Class Ib RNRs, in contrast, have a partial cone domain and have generally been found to be insensitive to dATP inhibition. An exception is the Bacillus subtilis Ib RNR, which we recently reported to be inhibited by physiological concentrations of dATP. Here, we demonstrate that the α subunit of this RNR contains tightly bound deoxyadenosine 5'-monophosphate (dAMP) in its N-terminal domain and that dATP inhibition of CDP reduction is enhanced by its presence. X-ray crystallography reveals a previously unobserved (noncanonical) α 2 dimer with its entire interface composed of the partial N-terminal cone domains, each binding a dAMP molecule. Using small-angle X-ray scattering (SAXS), we show that this noncanonical α 2 dimer is the predominant form of the dAMP-bound α in solution and further show that addition of dATP leads to the formation of larger oligomers. Based on this information, we propose a model to describe the mechanism by which the noncanonical α 2 inhibits the activity of the B. subtilis Ib RNR in a dATP- and dAMP-dependent manner.

59 BASIC BIOLOGICAL SCIENCES↗

Regulation of T cell expansion by antigen presentation dynamics

An essential feature of the adaptive immune system is the proliferation of antigen-specific lymphocytes during an immune reaction to form a large pool of effector cells. This proliferation must be regulated to ensure an effective response to infection while avoiding immunopathology. Recent experiments in mice have demonstrated that the expansion of a specific clone of T cells in response to cognate antigen obeys a striking inverse power law with respect to the initial number of T cells. Here, we show that such a relationship arises naturally from a model in which T cell expansion is limited by decaying levels of presented antigen. The same model also accounts for the observed dependence of T cell expansion on affinity for antigen and on the kinetics of antigen administration. Extending the model to address expansion of multiple T cell clones competing for antigen, we find that higher-affinity clones can suppress the proliferation of lower-affinity clones, thereby promoting the specificity of the response. Using the model to derive optimal vaccination protocols, we find that exponentially increasing antigen doses can achieve a nearly optimized response. We thus conclude that the dynamics of presented antigen is a key regulator of both the size and specificity of the adaptive immune response.

59 BASIC BIOLOGICAL SCIENCES↗

Divergent kinase regulates membrane ultrastructure of the Toxoplasma parasitophorous vacuole

Apicomplexan parasites replicate within a protective organelle, called the parasitophorous vacuole (PV). The Toxoplasma gondii PV is filled with a network of tubulated membranes, which are thought to facilitate trafficking of effectors and nutrients. Despite being critical to parasite virulence, there is scant mechanistic understanding of the network’s functions. Here, we identify the parasite-secreted kinase WNG1 (With-No-Gly-loop) as a critical regulator of tubular membrane biogenesis. WNG1 family members adopt an atypical protein kinase fold lacking the glycine rich ATPbinding loop that is required for catalysis in canonical kinases. Unexpectedly, we find that WNG1 is an active protein kinase that localizes to the PV lumen and phosphorylates PV-resident proteins, several ofwhich are essential for the formation of a functional intravacuolar network. Moreover, we show thatWNG1-dependent phosphorylation of these proteins is required for their membrane association, and thus their ability to tubulate membranes. Consequently, WNG1 knockout parasites have an aberrant PV membrane ultrastructure. Collectively, our results describe a unique family of Toxoplasma kinases and implicate phosphorylation of secreted proteins as a mechanism of regulating PV development during parasite infection.

59 BASIC BIOLOGICAL SCIENCES↗

Kv2.1 channels play opposing roles in regulating membrane potential, Ca 2+ channel function, and myogenic tone in arterial smooth muscle

The accepted role of the protein Kv2.1 in arterial smooth muscle cells is to form K + channels in the sarcolemma. Opening of Kv2.1 channels causes membrane hyperpolarization, which decreases the activity of L-type Ca V 1.2 channels, lowering intracellular Ca 2+ ([Ca 2+ ] i ) and causing smooth muscle relaxation. A limitation of this model is that it is based exclusively on data from male arterial myocytes. Here, we used a combination of electrophysiology as well as imaging approaches to investigate the role of Kv2.1 channels in male and female arterial myocytes. We confirmed that Kv2.1 plays a canonical conductive role but found it also has a structural role in arterial myocytes to enhance clustering of Ca V 1.2 channels. Less than 1% of Kv2.1 channels are conductive and induce membrane hyperpolarization. Paradoxically, by enhancing the structural clustering and probability of Ca V 1.2–Ca V 1.2 interactions within these clusters, Kv2.1 increases Ca 2+ influx. These functional impacts of Kv2.1 depend on its level of expression, which varies with sex. In female myocytes, where expression of Kv2.1 protein is higher than in male myocytes, Kv2.1 has conductive and structural roles. Female myocytes have larger Ca V 1.2 clusters, larger [Ca 2+ ] i , and larger myogenic tone than male myocytes. In contrast, in male myocytes, Kv2.1 channels regulate membrane potential but not Ca V 1.2 channel clustering. We propose a model in which Kv2.1 function varies with sex: in males, Kv2.1 channels control membrane potential but, in female myocytes, Kv2.1 plays dual electrical and Ca V 1.2 clustering roles. This contributes to sex-specific regulation of excitability, [Ca 2+ ] i , and myogenic tone in arterial myocytes.

59 BASIC BIOLOGICAL SCIENCES↗