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Reovirus Low-Density Particles Package Cellular RNA.

ABSTRACT: Packaging of segmented, double-stranded RNA viral genomes requires coordination of viral proteins and RNA segments. For mammalian orthoreovirus (reovirus), evidence suggests either all ten or zero viral RNA segments are simultaneously packaged in a highly coordinated process hypothesized to exclude host RNA. Accordingly, reovirus generates genome-containing virions and "genomeless" top component particles. Whether reovirus virions or top component particles package host RNA is unknown. To gain insight into reovirus packaging potential and mechanisms, we employed next-generation RNA-sequencing to define the RNA content of enriched reovirus particles. Reovirus virions exclusively packaged viral double-stranded RNA. In contrast, reovirus top component particles contained similar proportions but reduced amounts of viral double-stranded RNA and were selectively enriched for numerous host RNA species, especially short, non-polyadenylated transcripts. Host RNA selection was not dependent on RNA abundance in the cell, and specifically enriched host RNAs varied for two reovirus strains and were not selected solely by the viral RNA polymerase. Collectively, these findings indicate that genome packaging into reovirus virions is exquisitely selective, while incorporation of host RNAs into top component particles is differentially selective and may contribute to or result from inefficient viral RNA packaging.

SUBMITTER: Thoner TW

PROVIDER: S-EPMC8228547 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Reovirus low-density particles package viral and cellular RNA

Project description:Packaging of segmented, double-stranded RNA viral genomes requires coordination of multiple viral proteins and RNA segments. For mammalian orthoreovirus (reovirus), evidence suggests either all ten or zero viral RNA segments are simultaneously packaged in a highly coordinated process hypothesized to exclude host RNA. Accordingly, reovirus generates genome-containing virions and “genomeless” top component particles. However, despite ostensibly lacking the genome, top component particles maintain a low level of infectivity. Whether reovirus particles can package host RNA is unknown. To gain insight into reovirus packaging potential and mechanisms, we employed next-generation RNA-sequencing to define the viral and host RNA content of purified reovirus virions and top component particles. Reovirus top component particles contained double-stranded viral RNA segments in similar proportions but at reduced levels compared to virions. Top component particles also were enriched for numerous host RNAs, especially short, non-polyadenylated transcripts, that differed by reovirus strain, independent of the viral polymerase. In contrast, virions were enriched for very few host RNAs. Collectively, these findings indicate that genome packaging into reovirus virions is exquisitely selective, while incorporation of host RNAs into top component particles is more promiscuous or differentially selective and may contribute to or result from inefficient viral RNA packaging.

2021-06-08 | GSE164270 | GEO

Reovirus low-density particles package viral and cellular RNA

Project description:Reovirus low-density particles package viral and cellular RNA

| PRJNA689770 | ENA

Low-density hepatitis C virus infectious particles are protected from oxidation by secreted cellular proteins.

Project description:ImportanceAssessments of viral stability on surfaces or in body fluids under different environmental conditions and/or temperatures are often performed, as they are key to understanding the routes and parameters of viral transmission and to providing clues on the epidemiology of infections. However, for most viruses, the mechanisms of inactivation vs stability of viral particles remain poorly defined. Although they are structurally diverse, with different compositions, sizes, and shapes, enveloped viruses are generally less stable than non-enveloped viruses, pointing out the role of envelopes themselves in virus lability. In this report, we investigated the properties of hepatitis C virus (HCV) particles with regards to their stability. We found that, compared to alternative enveloped viruses such as Dengue virus (DENV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), hepatitis delta virus (HDV), and Crimean-Congo hemorrhagic fever virus (CCHFV) that infect the liver, HCV particles are intrinsically labile. We determined the mechanisms that drastically alter their specific infectivity through oxidation of their lipids, and we highlighted that they are protected from lipid oxidation by secreted cellular proteins, which can protect their membrane fusion capacity and overall infectivity.

| S-EPMC10653866 | biostudies-literature

Small dense low-density lipoprotein particles: clinically relevant?

Project description:Purpose of reviewLevels of small, dense low-density lipoprotein (LDL) (sdLDL) particles determined by several analytic procedures have been associated with risk of atherosclerotic cardiovascular disease (ASCVD). This review focuses on the clinical significance of sdLDL measurement.Recent findingsResults of multiple prospective studies have supported earlier evidence that higher levels of sdLDL are significantly associated with greater ASCVD risk, in many cases independent of other lipid and ASCVD risk factors as well as levels of larger LDL particles. A number of properties of sdLDL vs. larger LDL, including reduced LDL receptor affinity and prolonged plasma residence time as well as greater oxidative susceptibility and affinity for arterial proteoglycans, are consistent with their heightened atherogenic potential. Nevertheless, determination of the extent to which sdLDL can preferentially impact ASCVD risk compared with other apoprotein B-containing lipoproteins has been confounded by their metabolic interrelationships and statistical collinearity, as well as differences in analytic procedures and definitions of sdLDL.SummaryA growing body of data points to sdLDL concentration as a significant determinant of ASCVD risk. Although future studies should be aimed at determining the clinical benefit of reducing sdLDL levels, there is sufficient evidence to warrant consideration of sdLDL measurement in assessing and managing risk of cardiovascular disease.Video abstracthttps://www.dropbox.com/s/lioohr2ead7yx2p/zoom_0.mp4?dl=0.

| S-EPMC9197986 | biostudies-literature

Pollens derived magnetic porous particles for adsorption of low-density lipoprotein from plasma.

Project description:Adsorption of low-density lipoprotein from plasma is vital for the treatment of dyslipidemia. Appropriate adsorbent material for efficient and selective adsorption of low-density lipoprotein is highly desired. In this work, we developed pollens-derived magnetic porous particles as adsorbents for this purpose. The natural pollen grains were modified to obtain high surface porosity, a large inner cavity, magnet responsiveness, and specific wettability. The resultant particles exhibited satisfying performance in the adsorption of a series of oils and organic solvents out of water. Besides, the particles were directly utilized to the adsorption of low-density lipoprotein in plasma, which showed high selectivity, and achieved an outstanding adsorption capacity as high as 34.9% within 2 h. Moreover, their salient biocompatibility was demonstrated through simulative hemoperfusion experiments. These features, together with its abundant source and facile fabrication, makes the pollens-derived magnetic porous particles excellent candidate for low-density lipoprotein -apheresis and water treatment applications.

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High hydrostatic pressure specifically affects molecular dynamics and shape of low-density lipoprotein particles.

Project description:Lipid composition of human low-density lipoprotein (LDL) and its physicochemical characteristics are relevant for proper functioning of lipid transport in the blood circulation. To explore dynamical and structural features of LDL particles with either a normal or a triglyceride-rich lipid composition we combined coherent and incoherent neutron scattering methods. The investigations were carried out under high hydrostatic pressure (HHP), which is a versatile tool to study the physicochemical behavior of biomolecules in solution at a molecular level. Within both neutron techniques we applied HHP to probe the shape and degree of freedom of the possible motions (within the time windows of 15 and 100?ps) and consequently the flexibility of LDL particles. We found that HHP does not change the types of motion in LDL, but influences the portion of motions participating. Contrary to our assumption that lipoprotein particles, like membranes, are highly sensitive to pressure we determined that LDL copes surprisingly well with high pressure conditions, although the lipid composition, particularly the triglyceride content of the particles, impacts the molecular dynamics and shape arrangement of LDL under pressure.

| S-EPMC5382586 | biostudies-literature

Characterization of hepatitis C RNA-containing particles from human liver by density and size.

Project description:Hepatitis C virus (HCV) particles found in vivo are heterogeneous in density and size, but their detailed characterization has been restricted by the low titre of HCV in human serum. Previously, our group has found that HCV circulates in blood in association with very-low-density lipoprotein (VLDL). Our aim in this study was to characterize HCV RNA-containing membranes and particles in human liver by both density and size and to identify the subcellular compartment(s) where the association with VLDL occurs. HCV was purified by density using iodixanol gradients and by size using gel filtration. Both positive-strand HCV RNA (present in virus particles) and negative-strand HCV RNA (an intermediate in virus replication) were found with densities below 1.08 g ml(-1). Viral structural and non-structural proteins, host proteins ApoB, ApoE and caveolin-2, as well as cholesterol, triglyceride and phospholipids were also detected in these low density fractions. After fractionation by size with Superose gel filtration, HCV RNA and viral proteins co-fractionated with endoplasmic reticulum proteins and VLDL. Fractionation on Toyopearl, which separates particles with diameters up to 200 nm, showed that 78 % of HCV RNA from liver was >100 nm in size, with a positive-/negative-strand ratio of 6 : 1. Also, 8 % of HCV RNA was found in particles with diameters between 40 nm and 70 nm and a positive-/negative-strand ratio of 45 : 1. This HCV was associated with ApoB, ApoE and viral glycoprotein E2, similar to viral particles circulating in serum. Our results indicate that the association between HCV and VLDL occurs in the liver.

| S-EPMC2557069 | biostudies-literature

Conformational changes accompany activation of reovirus RNA-dependent RNA transcription.

Project description:Many critical biologic processes involve dynamic interactions between proteins and nucleic acids. Such dynamic processes are often difficult to delineate by conventional static methods. For example, while a variety of nucleic acid polymerase structures have been determined at atomic resolution, the details of how some multi-protein transcriptase complexes actively produce mRNA, as well as conformational changes associated with activation of such complexes, remain poorly understood. The mammalian reovirus innermost capsid (core) manifests all enzymatic activities necessary to produce mRNA from each of the 10 encased double-stranded RNA genes. We used rapid freezing and electron cryo-microscopy to trap and visualize transcriptionally active reovirus core particles and compared them to inactive core images. Rod-like density centered within actively transcribing core spike channels was attributed to exiting nascent mRNA. Comparative radial density plots of active and inactive core particles identified several structural changes in both internal and external regions of the icosahedral core capsid. Inactive and transcriptionally active cores were partially digested with trypsin and identities of initial tryptic peptides determined by mass spectrometry. Differentially-digested peptides, which also suggest transcription-associated conformational changes, were placed within the known three-dimensional structures of major core proteins.

| S-EPMC2819412 | biostudies-literature

Lowering Low-Density Lipoprotein Particles in Plasma Using Dextran Sulphate Co-Precipitates Procoagulant Extracellular Vesicles.

Project description:Plasma extracellular vesicles (EVs) are lipid membrane vesicles involved in several biological processes including coagulation. Both coagulation and lipid metabolism are strongly associated with cardiovascular events. Lowering very-low- and low-density lipoprotein ((V)LDL) particles via dextran sulphate LDL apheresis also removes coagulation proteins. It remains unknown, however, how coagulation proteins are removed in apheresis. We hypothesize that plasma EVs that contain high levels of coagulation proteins are concomitantly removed with (V)LDL particles by dextran sulphate apheresis. For this, we precipitated (V)LDL particles from human plasma with dextran sulphate and analyzed the abundance of coagulation proteins and EVs in the precipitate. Coagulation pathway proteins, as demonstrated by proteomics and a bead-based immunoassay, were over-represented in the (V)LDL precipitate. In this precipitate, both bilayer EVs and monolayer (V)LDL particles were observed by electron microscopy. Separation of EVs from (V)LDL particles using density gradient centrifugation revealed that almost all coagulation proteins were present in the EVs and not in the (V)LDL particles. These EVs also showed a strong procoagulant activity. Our study suggests that dextran sulphate used in LDL apheresis may remove procoagulant EVs concomitantly with (V)LDL particles, leading to a loss of coagulation proteins from the blood.

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Migration Behavior of Low-Density Particles in Lab-on-a-Disc Devices: Effect of Walls.

Project description:The effect of the lateral walls of a Lab-On-a-Disc device on the dynamics of a model system of particles with a density lower than that of the solvent (modelling parasites eggs) is analyzed theoretically and experimentally. In the absence of lateral walls, a particle always moves in the direction of the centrifugal force, while its trajectory is deflected in the tangential direction by the inertial Coriolis and Euler forces. Lateral walls, depending on the angle forming with the radial direction, can guide the particle either in the same or in the opposite direction to the centrifugal force, thus resulting in unusual particle trajectories including zig-zag or backwards particle motion. The effect is pronounced in the case of short operation times when the acceleration of the angular rotation, and thus the Euler force, is considerable. The predicted unusual motion is demonstrated by numerically solving the equation of motion in the presence of lateral walls and verified in the experiment with particles of density lower than that of the solvent. Our analysis is useful for design and operational considerations of Lab-On-a-Disc devices aiming for or involving (bio)particle handling.

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