Nucleosome Research Articles

Structural roles of Ump1 and β-subunit propeptides in proteasome biogenesis.

The yeast pre1-1(β4-S142F) mutant accumulates late 20S proteasome core particle precursor complexes (late-PCs). We report a 2.1 Å cryo-EM structure of this intermediate with full-length Ump1 trapped inside, and Pba1-Pba2 attached to the α-ring surfaces. The structure discloses intimate interactions of Ump1 with β2- and β5-propeptides, which together fill most of the antechambers between the α- and β-rings. The β5-propeptide is unprocessed and separates Ump1 from β6 and β7. The β2-propeptide is disconnected from the subunit by autocatalytic processing and localizes between Ump1 and β3. A comparison of different proteasome maturation states reveals that maturation goes along with global conformational changes in the rings, initiated by structuring of the proteolytic sites and their autocatalytic activation. In the pre1-1 strain, β2 is activated first enabling processing of β1-, β6-, and β7-propeptides. Subsequent maturation of β5 and β1 precedes degradation of Ump1, tightening of the complex, and finally release of Pba1-Pba2.

Structural basis for human OGG1 processing 8-oxodGuo within nucleosome core particles

Base excision repair (BER) is initialized by DNA glycosylases, which recognize and flip damaged bases out of the DNA duplex into the enzymes active site, followed by cleavage of the glycosidic bond. Recent studies have revealed that all types of DNA glycosylases repair base lesions less efficiently within nucleosomes, and their repair activity is highly depended on the lesion’s location within the nucleosome. To reveal the underlying molecular mechanism of this phenomenon, we determine the 3.1 Å cryo-EM structure of human 8-oxoguanine-DNA glycosylase 1 (hOGG1) bound to a nucleosome core particle (NCP) containing a common oxidative base lesion, 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo). Our structural analysis shows that hOGG1 can recognize and flip 8-oxodGuo even within NCPs; however, the interaction between 8-oxodGuo and hOGG1 in a NCP context is weaker than in free DNA due to competition for nucleosomal DNA by the histones. Binding of OGG1 and the flipping of 8-oxodGuo by hOGG1 leads to a partial detachment of DNA from the histone core and a ratchet-like inward movement of nucleosomal DNA. Our findings provide insights into how the dynamic structure of nucleosomes modulate the activity of repair enzymes within chromatin.

Design and damping characterization of sandwich composite made of particle-filled hollow spheres and steel sheets

Sandwich composites made of particle-filled hollow sphere structures (PHSS) and steel sheets offer excellent lightweight and damping properties, ideal for high-speed machine tool components under dynamic loads. Previous research has focused on single particle-filled hollow sphere (PHS) and small PHSS, leaving a gap understanding of PHSS/steel sandwich composites. In this study a test rig was developed, and Design of Experiments and Response Surface Analysis were used to investigate the effects of sheet and core thickness (design parameters), filling ratio, and particle size (particle parameters) on the damping performance of PHSS/steel sandwiches. The results indicate that the design parameters have a significant influence on damping performance. The interaction between design and particle parameters also substantially affects damping. Minimizing particle size, increasing filling ratio, thinning the face layer, and thickening the core layer significantly improve structural damping. To address manufacturing tolerances, a finite element (FE) model-based optimization was developed to accurately determine PHSS material parameters. These parameters were used in an FE model of the PHSS-steel composite, with identified contact parameters minimizing measurement and simulation differences. The homogenized material model and the linear model using global damping parameters accurately reproduce the dynamic properties of the PHSS-steel sandwich composite in low vibration modes.

Facile Preparation of Raspberry-Like SiO2@Polyurea Microspheres with Tunable Wettability and Their Application for Oil-Water Separation.

Raspberry-like microspheres have been widely used as superhydrophobic materials, photonic crystals, drug carriers, etc. Nevertheless, their preparation methods, usually consisting of multiple steps, are generally time- and energy-consuming. Herein raspberry-like SiO2@polyurea microspheres (SiO2@PUM) are readily prepared via a one-step precipitation polymerization of isophorone diisocyanate in a H2O/acetone mixture with the presence of SiO2 particles. The sphere size, surface roughness, and SiO2 content of SiO2@PUM are easily adjustable by varying the experimental conditions. TEM and SEM observations reveal that the final SiO2@PUM exhibits a core-shell structure, with polyurea (PU) in the core and SiO2 particles as the shell. In the process, the SiO2 particles were initially located on the PUM surface as a monolayer. With the reaction proceeding, the monolayer of SiO2 particles became thicker, forming a thicker layer of SiO2 particles on PUM due to the accumulation of SiO2 particles, leading to a multilayer structure of SiO2 particles on the shell of SiO2@PUM. The formation mechanism of the raspberry-like SiO2@PUM was thoroughly discussed and ascribed to electrostatic attraction between the positively charged PU and negatively charged SiO2 particles. Once dried, SiO2@PUM was superhydrophobic and turned hydrophilic if water-wetted. Using a layer of SiO2@PUM, effective separation with good reusability for a variety of oil-water mixtures was achieved regardless of the oil density and types of oil-water emulsions. This work presents a novel protocol for the preparation of raspberry-like microspheres with tunable wettability via a rapid and green process, and the resulting microspheres are highly effective for the separation of diverse types of oil-water mixtures.

Probing the Nanonewton Mitotic Cell Deformation Force by Ion-Resonance-Enhanced Photonics Force Microscopy.

Mechanical forces are essential for regulating dynamic changes in cellular activities. A comprehensive understanding of these forces is imperative for unraveling fundamental mechanisms. Here, we develop a microprobe capable of facilitating the measurement of biological forces up to nanonewton levels in living cells. This probe is designed by coating the core of anatase titania particles with amorphous titania and silica shells and an upconversion nanoparticles (UCNPs) layer. Leveraging both antireflection and ion resonance effects from the shells, the optically trapped probe attains a maximum lateral optical trap stiffness of 14.24 pN μm-1 mW-1, surpassing the best reported value by a factor of 3. Employing this advanced probe in a photonic force microscope, we determine the elasticity modulus of mitotic HeLa cells as 1.27 ± 0.3 kPa. Nanonewton probes offer the potential to explore 3D cellular mechanics with unparalleled precision and spatial resolution, fostering a deeper understanding of the underlying biomechanical mechanisms.

Ten questions concerning particle exposure in open electrosurgery and its control strategies

The increasing use of high-frequency electrosurgical pencils in open surgeries has heightened concerns about the adverse implications of electrosurgical smoke (ES) on perioperative personnel. Since 2015, there has been an increasing amount of research on electrosurgical smoke particles annually. Despite data discrepancies, nanoparticles (core particle size <200 nm) generally dominate in number distributions. Particulate matter (PM) concentration is closely related to the type of target tissue. Most studies show that ultrafine particle (UFP) concentrations near the main surgeon’s breathing zone inversely correlate with operating room air changes and positively correlate with the power of the electrosurgical pencil. Local smoke evacuation devices with negative ions, specialized filters and N95 respirators are more effective in protecting perioperative staff health, particularly against UFPs. The objective is to summarize particle exposure knowledge in open electrosurgery and establish a research agenda to protect medical personnel from ES exposure.

Experimental study on the rock breaking effect of different tooth shapes of inserted teeth in a hobbing-cone hybrid PDC bit under complex stress environment

The hobbing-cone hybrid PDC bit has emerged with the increasing exploitation of deep oil and gas resources. As an important component of new bits, studying the mechanism of toothed fracturing of deep rocks is crucial for improving rock-breaking efficiency. In this paper, the spherical teeth and conical teeth were used to intrude sandstone under different confining pressures, and the brittleness index (BIm), specific energy (SE), and average fragment size (dm) were used to analyze the rock-breaking effect. The 3D contour scanner quantitatively analyzed the crushing pit area. Finally, the in-situ CT scanning and discrete element numerical simulation were used to summarize the crack propagation law. The results show that with the increase of confining pressure, the BIm of inserted teeth increases continuously, and the growth rate of conical teeth increases, while that of spherical teeth decreases. The SE and dm exhibit a symmetrical distribution with a confining pressure of 10MPa as the axis of symmetry. When the difference in principal stress is 5MPa, the SE is the lowest point, the rock-breaking efficiency is the highest, and the corresponding dm is at the highest point, but the curve trend is the opposite. When the stress difference is 5MPa, it is conducive to propagating surface cracks, thus forming a larger crushing area and the highest rock-breaking efficiency. Based on CT results, the dense particle core is formed near the inserted teeth, and that of the spherical teeth is located below. In contrast, that of the conical teeth is located on the side, which is also the reason for the difference in crack propagation between different tooth shapes.

Creating a bacterium that forms eukaryotic nucleosome core particles

The nucleosome is one of the hallmarks of eukaryotes, a dynamic platform that supports many critical functions in eukaryotic cells. Here, we engineer the in vivo assembly of the nucleosome core in the model bacterium Escherichia coli. We show that bacterial chromosome DNA and eukaryotic histones can assemble in vivo to form nucleosome complexes with many features resembling those found in eukaryotes. The formation of nucleosomes in E. coli was visualized with atomic force microscopy and using tripartite split green fluorescent protein. Under a condition that moderate histones expression was induced at 1 µM IPTG, the nucleosome-forming bacterium is viable and has sustained growth for at least 110 divisions in longer-term growth experiments. It exhibits stable nucleosome formation, a consistent transcriptome across passages, and reduced growth fitness under stress conditions. In particular, the nucleosome arrays in E. coli genic regions have profiles resembling those in eukaryotic cells. The observed compatibility between the eukaryotic nucleosome and the bacterial chromosome machinery may reflect a prerequisite for bacteria-archaea union, providing insight into eukaryogenesis and the origin of the nucleosome.

Λ6He (1−, 2−) spin doublet in a Λ + n + α cluster model

The spin singlet and spin triplet states of the helium-6-lambda hypernucleus are studied in configuration space as a three-body problem, using the hyperspherical method. This hypernucleus is treated as a Λ + n + α cluster model. Since the α particle core has a spin of 0, this is an ideal hypernucleus from which to understand spin dependence in hyperon-nucleon potentials. In this paper, the hyperon-nucleon potential considered is the GLM-YN0 Λ-n potential, which is a simulation of the NSC97f potential obtained through inverse scattering theory. Due to violations of the cluster structure, two-body potentials underbind the helium-6-lambda hypernucleus. This issue is addressed by including a three-body force to complement these two-body forces. A three-body force, whose strength is spin dependent, is used to guide predictions of the spin triplet state of helium-6-lambda which is not yet known experimentally.

Electrostatic interactions between soft nanoparticles beyond the Derjaguin approximation: Effects of finite size of ions and charges, dielectric decrement and ion correlations

HypothesisElectrostatic interactions between colloids are governed by the overlap of their electric double layers (EDLs) and the ionic screening of the structural charges distributed at their core surface and/or in their peripheral ion-permeable shell, relevant to soft particles like polymer colloids and microorganisms. Whereas ion size-mediated effects on the organization of isolated EDLs have been analysed, their contribution to the electrostatic energy of interacting soft particles has received less attention Theory and simulationsHerein, we elaborate a formalism to evaluate the electrostatic interaction energy profile between spherical core/shell particles, building upon a recent Poisson-Boltzmann theory corrected for the sizes of ions and particle structural charges, for ion correlations and dielectric decrement. Interaction energy is derived from pairwise disjoining pressure and exact Surface Element Integration method, beyond the Derjaguin approximation. The theory is sufficiently flexible to tackle homo- and hetero-interactions that involve weakly to highly charged hard, porous or core/shell nano- to micro-sized particles in asymmetric multivalent electrolytes. FindingsResults illustrate how ion steric effects, ion correlations and dielectric decrement impact the sign, magnitude and range of the interactions depending on the particle size, the Debye length, and the geometric and electrostatic properties of the particle core and shell components.

CDCA7 is an evolutionarily conserved hemimethylated DNA sensor in eukaryotes.

Mutations of the SNF2 family ATPase HELLS and its activator CDCA7 cause immunodeficiency, centromeric instability, and facial anomalies syndrome, characterized by DNA hypomethylation at heterochromatin. It remains unclear why CDCA7-HELLS is the sole nucleosome remodeling complex whose deficiency abrogates the maintenance of DNA methylation. We here identify the unique zinc-finger domain of CDCA7 as an evolutionarily conserved hemimethylation-sensing zinc finger (HMZF) domain. Cryo-electron microscopy structural analysis of the CDCA7-nucleosome complex reveals that the HMZF domain can recognize hemimethylated CpG in the outward-facing DNA major groove within the nucleosome core particle, whereas UHRF1, the critical activator of the maintenance methyltransferase DNMT1, cannot. CDCA7 recruits HELLS to hemimethylated chromatin and facilitates UHRF1-mediated H3 ubiquitylation associated with replication-uncoupled maintenance DNA methylation. We propose that the CDCA7-HELLS nucleosome remodeling complex assists the maintenance of DNA methylation on chromatin by sensing hemimethylated CpG that is otherwise inaccessible to UHRF1 and DNMT1.

Structure of anellovirus-like particles reveal a mechanism for immune evasion

Anelloviruses are nonpathogenic viruses that comprise a major portion of the human virome. Despite being ubiquitous in the human population, anelloviruses (ANVs) remain poorly understood. Basic features of the virus, such as the identity of its capsid protein and the structure of the viral particle, have been unclear until now. Here, we use cryogenic electron microscopy to describe the first structure of an ANV-like particle. The particle, formed by 60 jelly roll domain-containing ANV capsid proteins, forms an icosahedral particle core from which spike domains extend to form a salient part of the particle surface. The spike domains come together around the 5-fold symmetry axis to form crown-like features. The base of the spike domain, the P1 subdomain, shares some sequence conservation between ANV strains while a hypervariable region, forming the P2 subdomain, is at the spike domain apex. We propose that this structure renders the particle less susceptible to antibody neutralization by hiding vulnerable conserved domains while exposing highly diverse epitopes as immunological decoys, thereby contributing to the immune evasion properties of anelloviruses. These results shed light on the structure of anelloviruses and provide a framework to understand their interactions with the immune system.

Citrate stabilized maghemite hydrosol with controllable MRI contrast: Key role of nanoparticle size

In this study, we have synthetized a series of citric acid stabilized superparamagnetic iron oxide nanoparticles (CA-SPIONs) with different core sizes in an automated chemical reactor with high repeatability of the nanoparticle size and chemical composition. The prepared CA-SPIONs are highly crystalline spherical-shaped particles with the diameters of 3.5 ± 0.7, 6 ± 1, 9 ± 1, and 12 ± 2 nm. The valent state of iron oxide was determined by a combination of X-ray photoelectron spectroscopy, UV–Vis spectroscopy, and Mössbauer studies, which confirmed predominantly maghemite formation. Under normal conditions, these nanoparticles exhibit no coercive force and no hysteresis, while saturation magnetization increases from 2 to 61 emu/g along with the increasing core size. Both longitudinal (r1) and transverse (r2) relaxivities of maghemite hydrosols with different nanoparticle sizes were measured and compared with the same data for the commercial Gd-complex (Gadovist). Magnetic circular dichroism spectroscopy indicated that aggregation occurs in magnetic field, but 9 nm samples slightly aggregate in the fields above 1.0 T, whereas 3.5 nm colloids are stable and do not exhibit aggregation behavior even at 1.5 T. The obtained series were examined in phantom test in clinical 1.5 T MRI scanner, which showed that increasing the particle core size resulted in an enhanced T2 contrast, while T1 contrast declined. Finally, the smallest CA-SPION colloid nanoparticles with the size of 3.5 nm exhibited significant T1 contrast enhancement, comparable with the commercial Gd-complex in water and human plasma as well. The maghemite hydrosol formed by nanoparticles with 3.5 nm size thus has a promising future as a T1 MRI contrast agent.

High‐Load Core@Shell Nanocarriers with Irinotecan and 5‐Fluorouracil for Combination Chemotherapy in Colorectal Cancer

Colorectal cancer (CRC) is the third most common cancer type and second leading cause of cancer‐related deaths worldwide, requiring novel drug‐delivery concepts. ITC@ZrO(TocP)/ZrO(FdUMP) core@shell nanocarriers (designated ITC‐FdUMP‐NC) with the clinically relevant chemotherapeutics irinotecan (ITC) and fluoro‐2′‐deoxyuridine‐5′‐phosphate (FdUMP) (active derivative of 5′‐fluorouracil/5‐FU) are a new type of nanocarrier with high drug payload (22 wt% of lipophilic ITC: particle core; 10 wt% of hydrophilic FdUMP: particle shell). The nanocarriers are tested in different CRC cell lines, a normal cell line, and rectal cancer patient‐derived organoids (PDOs). Fluorescence‐labeled nanocarriers show efficient uptake by all CRC cells and allow to distinctly track the intracellular trafficking toward endolysosomal compartments. Although free chemotherapeutic drugs exhibit a greater potency in 2D cell cultures, ITC‐FdUMP‐NC demonstrate equivalent cytotoxic efficacies as the freely dissolved drugs in the more complex 3D rectal cancer PDOs. The sustained drug‐release profile of the nanocarriers contrasts favorably with conventional free drugs, potentially enhancing the therapeutic outcome in vivo. With a chemotherapeutic cocktail comparable to the clinically applied FOLFIRI (ITC + 5‐FU), the ITC‐FdUMP‐NC represent a novel type of nanocarrier with high anti‐tumor effect and high drug payload, offering a promising strategy to circumvent chemoresistance and to improve therapy efficacy in vivo with less side effects.

DIII-D research to provide solutions for ITER and fusion energy

The DIII-D tokamak has elucidated crucial physics and developed projectable solutions for ITER and fusion power plants in the key areas of core performance, boundary heat and particle transport, and integrated scenario operation, with closing the core-edge integration knowledge gap being the overarching mission. New experimental validation of high-fidelity, multi-channel, non-linear gyrokinetic turbulent transport models for ITER provides strong confidence it will achieve Q ⩾ 10 operation. Experiments identify options for easing H-mode access in hydrogen, and give new insight into the isotopic dependence of transport and confinement. Analysis of 2,1 islands in unoptimized low-torque IBS demonstration discharges suggests their onset time occurs randomly in the constant β phase, most often triggered by non-linear 3-wave coupling, thus identifying an NTM seeding mechanism to avoid. Pure deuterium SPI for disruption mitigation is shown to provide favorable slow cooling, but poor core assimilation, suggesting paths for improved SPI on ITER. At the boundary, measured neutral density and ionization source fluxes are strongly poloidally asymmetric, implying a 2D treatment is needed to model pedestal fuelling. Detailed measurements of pedestal and SOL quantities and impurity charge state radiation in detached divertors has validated edge fluid modelling and new self-consistent ‘pedestal-to-divertor’ integrated modeling that can be used to optimize reactors. New feedback adaptive ELM control minimizes confinement reduction, and RMP ELM suppression with sustained high core performance was obtained for the first time with the outer strike point in a W-coated, compact and unpumped small-angle slot divertor. Advances have been made in integrated operational scenarios for ITER and power plants. Wide pedestal intrinsically ELM-free QH-modes are produced with more reactor-relevant conditions, Low torque IBS with W-equivalent radiators can exhibit predator-prey oscillations in T e and radiation which need control. High-β P scenarios with q min > 2, q 95–7.9, β N > 4, β T–3.3% and H 98y2 > 1.5 are sustained with high density ( n¯ = 7E19 m−3, f G–1) for 6 τ E, improving confidence in steady-state tokamak reactors. Diverted NT plasmas achieve high core performance with a non-ELMing edge, offering a possible highly attractive core-edge integration solution for reactors.

Atomistic Insights into Impact-Induced Energy Release and Deformation of Core-Shell-Structured Ni/Al Nanoparticle in an Oxygen Environment.

In actual atmospheric environments, Ni/Al composites subjected to high-velocity impact will undergo both intermetallic reaction and oxidative combustion simultaneously, and the coupling of mechanical and multiple chemical processes leads to extremely complex characteristics of energy release. This work employs ReaxFF molecular dynamics simulations to investigate the impact-induced deformation and energy release of a core-shell-structured Ni/Al nanoparticle in an oxygen environment. It was found that Al directly undergoes fragmentation, while Ni experiences plastic deformation, melting, and fragmentation in sequence as the impact velocity increased. This results in the final morphology of the nanoparticles being an ellipsoidal-clad nanoparticle, spherical Ni/Al melt, and debris cloud. Furthermore, these deformation characteristics are strongly related to the material property of the shell, manifested as Ni shell-Al core particle, being more prone to breakage. Interestingly, the dissociation phenomenon of Ni-Al-O clusters during deformation is observed, which is driven by Ni dissociation and Al oxidation. In addition, the energy release is strongly related to the deformation behavior. When the nanoparticle is not completely broken (Ni undergoes plastic deformation and melting), the energy release comes from the oxidative combustion of Al fragments and the intermetallic reaction driven by atomic mixing. When the nanoparticle is completely broken, the energy release mainly comes from the oxidative combustion of the debris cloud. At the same time, the promoting effect of oxygen concentration on the energy release efficiency is examined. These findings can provide atomic insights into the regulation of impact-induced energy release for reactive intermetallic materials.

(Invited) Plasmonic Yolk@Shell Nanocrystals for Photocatalytic Hydrogen Production

Plasmonic photocatalysts have garnered significant attention for hydrogen production due to their unique properties and efficiency in harnessing solar energy. This work is designed to develop yolk@shell nanocrystals as a high-efficiency photocatalyst model. The core yolk is composed of Au nanoparticles, and the shell is composed of hollow non-stoichiometric semiconductors. The compositional defects can induce a large number of free carriers, making non-stoichiometric semiconductors behave like noble metals and exhibit surface plasmon resonance (SPR) absorption. For yolk@shell nanocrystals, the core Au particles can move freely, which can effectively stir the surrounding reaction solution to increase the reaction rate. The confined space between the core particle and the shell layer can function as a nanoreactor to promote species diffusion to enhance catalysis kinetics. In addition, the core Au particles and the shell semiconductor can form metal/semiconductor heterojunction so as to exhibit remarkable photocatalytic performance. Several representative works from our lab will be introduced to demonstrate the promising potentials of plasmonic yolk@shell nanocrystals for hydrogen production applications.

Design of Modular, 3D‐Printed Millifluidic Mixers to Enable Sequential NanoPrecipitation (SNaP) for the Tunable Synthesis of Drug‐Loaded Nanoparticles and Microparticles

AbstractSequential NanoPrecipitation (SNaP) is a nascent controlled precipitation process for the tunable formation of polymeric particles for drug delivery and bioimaging. While SNaP utilizes the same self‐assembly principles as one‐step Flash NanoPrecipitation, SNaP is a two‐step assembly process in which the particle core formation is initiated during a first mixing step followed by particle stabilization in a second mixing step. Current SNaP experimental set‐ups use commercial millifluidic mixers connected in series, which have several limitations, including the inability to access short inter‐mixer delay times (Td). A robust, 3D‐printed, modular mixer design that enables access to short Td's (〈 25 ms) not previously accessible is reported. For the first time, it is demonstrated that decoupling the assembly steps improves control over particle size, expanding the attainable size range to include both nanoparticles and microparticles. It is empirically proven that inter‐mixer Td is a key parameter for particle size control and that particle size scales with Td in agreement with Smoluchowski's model of diffusion‐limited growth. The formation of particles ranging in size from 160 nm to 1.2 µm is shown. Finally, the applicability of the new mixers is established by encapsulating fluorophores and therapeutics into particles for the first time via SNaP.

Axisymmetric Slow Rotation of Coaxial Soft/Porous Spheres.

The steady low-Reynolds-number rotation of a chain of coaxial soft spheres (each with an impermeable hard core covered by a permeable porous layer) about the axis in a viscous fluid is analyzed. The particles may be unequally spaced, and may differ in the permeability and inner and outer radii of the porous surface layer as well as angular velocity. By using a method of boundary collocation, the Stokes and Brinkman equations for the external fluid flow and flow within the surface layers, respectively, are solved semi-analytically. The particle interaction effect increases as the relative gap thickness between adjacent particles or their permeability decreases, which can be significant as the gap thickness approaches zero. A particle's hydrodynamic torque is reduced (its rotation is enhanced) when other particles rotate in the same direction at equivalent or greater angular velocities, but increases (its rotation is hindered) when other particles rotate in the opposite direction at arbitrary angular velocities. For particles with different radii or permeabilities, the particle interaction has a greater effect on smaller or more permeable particles than on larger or less permeable particles. For the rotation of three particles, the presence of the third particle can significantly affect the hydrodynamic torques acting on the other two particles. For the rotation of numerous particles, shielding effects between particles can be substantial. When the permeability of porous layers is low, relative fluid motion is barely felt by the hard cores of the soft particles. The insights gained from this analysis on the effects of interactions among rotating soft particles may be of great importance in many physicochemical applications of colloidal suspensions.

ILP1 and NTR1 affect the stability of U6 snRNA during spliceosome complex disassembly in Arabidopsis

U6 snRNA is one of the uridine-rich non-coding RNAs, abundant and stable in various cells, function as core particles in the intron-lariat spliceosome (ILS) complex. The Increased Level of Polyploidy1–1D (ILP1) and NTC-related protein 1 (NTR1), two conserved disassembly factors of the ILS complex, facilitates the disintegration of the ILS complex after completing intron splicing. The functional impairment of ILP1 and NTR1 lead to increased U6 levels, while other snRNAs comprising the ILS complex remained unaffected. We revealed that ILP1 and NTR1 had no impact on the transcription, 3’ end phosphate structure or oligo(U) tail of U6 snRNA. Moreover, we uncovered that the mutation of ILP1 and NTR1 resulted in the accumulation of ILS complexes, impeding the dissociation of U6 from splicing factors, leading to an extended half-life of U6 and ultimately causing an elevation in U6 snRNA levels. Our findings broaden the understanding of the functions of ILS disassembly factors ILP1 and NTR1, and providing insights into the dynamic disassembly between U6 and ILS.