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Anderson, P.

Publications and source records attributed to Anderson, P..

At least 19 recordsLinked to original sources

A Stickiness Response System in Rats

The perception of stickiness is known to everyone who interacts with the world. While eating, walking, and navigating diverse environments including crowded subways, forests, fields and lunchrooms, stickiness is a common and old sensation. Responses to sticky stimuli have been measured in animal and human brains; however, precise behavioral responses and the underlying neural mechanisms are not well understood. We applied sticky stimuli to three-week-old rat pups and found the effects vary greatly across the animals body: Sticky stimuli are quickly removed from forepaws and nose, but often evoke only little reaction from hindpaws. When we applied sticky (marshmallow, mochi) and non-sticky stimuli (water, oil) to forepaws, we observed stimulus unspecific behaviors (licking and grooming) with variable response onsets as well as three fast-onset sticky-specific behaviors. Sticky-specific behaviors were exclusively triggered by sticky stimuli and included paw shaking and paw swiping (behaviors presumably aiming at stickiness removal) and paw tapping. In tapping, animals gently tap their forepaws onto each other or on the ground; we wondered if the resulting paw-substrate detachments serve stickiness sensing. Blocking of forepaw skin sensation reduced sticky-specific responses to sticky stimuli compared to control conditions (Ringer injections). To assess central representations of stickiness, we obtained in vivo whole-cell recordings of neurons in forepaw-somatosensory-cortex while presenting sticky and non-sticky stimuli to anesthetized rat pups. While responses were heterogeneous across the population, we observed individual neurons that had significantly different responses to stimulus detachment for sticky and non-sticky stimuli. In summary, we describe a fast-onset, body-part-specific stickiness response system in rats, which is strongly driven by forepaw skin afferents.

neuroscience↗

A uniform tissue-clearing framework and mesoSPIM-ultra enable cm-scale single-neuron tracing

Tissue-clearing and light-sheet microscopy have transformed volumetric imaging of intact organs, yet limited mechanistic understanding of dehydration-based clearing continues to constrain rational protocol design and broader applicability. Here, we define the cardinal chemical and physical principles underlying dehydration-based tissue-clearing and establish a new pipeline for large-volume imaging. To maximize imaging performance, we developed the "mesoSPIM-ultra", an upgraded mesoSPIM platform with a temperature-controlled sample chamber, a large field-of-view (FoV) camera and specialized optics to achieve long-working-distance, high-resolution imaging of cleared samples. We applied this approach to investigate the projectome of Chx10+ neurons, a cell population with complex axonal morphologies along the entire mouse spinal-cord and brain, and implicated in ipsilateral orienting behaviors. By combining behavioral analysis with post-hoc single-neuron reconstructions, we revealed previously inaccessible branching architectures and long-range projections extending from the brainstem to the spinal cord. Together, our work establishes a mechanistic foundation for tissue-clearing and scalable imaging.

neuroscience↗

A rapid, sensitive, and quantitative high plex biomarker digital detection platform enabled by Hypercoding

Low-cost, multiplexed, and automated assays are needed to make omic technologies more broadly accessible in clinical, research, and commercial settings. We present Hypercoding, a scalable technology for detection and quantitation of multi-omic targets. Drawing from data reliability methods in the telecommunications field, Hypercoding uses fluorescent signals from hybridization with an error-correcting code to enable detection of high-plexity targets from biological samples, such as human DNA. In the presence of the target, a linear DNA construct is circularized, immobilized, and amplified to enable single-molecule detection of a target via rapid readout cycles within a 96-well plate. We demonstrate capability for >10,000 code plexity and accurate (98.7%) genotyping of 209 pharmacogenomic variants. Furthermore, we show computation of copy number variation with whole chromosome and sub-gene resolution, as well as quantitation of target abundance down to 10 fM sensitivity with a dynamic range of up to 10 logs.

genomics↗

Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation

Emerging evidence strongly supports a close relationship between age-related hearing loss and frailty, highlighting the importance of early detection and intervention. Recently, we invented a mitochondria-homing drug named mitochonic acid 5 (MA-5), that increases the adenosine triphosphate (ATP) levels, rescue mitochondrial function, and protect tissue damages. Currently, the phase I clinical trial has been finished in Japan (jRCT2031210495) and the phase 2 clinical trial has already been approved by PMDA. Here we show that MA-5 improved various types of hearing loss in mouse models. Structural chemical bioanalysis revealed that MA-5 is a mixture of equal amount of S- and R- enantiomer and both S- and R- enantiomer increase ATP by binding mitochondrial protein, mitofilin. However, S-enantiomer significantly increased the NAD+ levels by binding to the NAD+-producing key enzyme nicotinamide phosphoribosyltransferase (NAMPT). Moreover, the S-enantiomer increased the sirtuin 1 protein by suppressing polyubiquitination induced by tripartite motif containing 28 (TRIM28) phosphorylation which was triggered by DNA-dependent protein kinase (DNA-PK) activation in the absence of DNA damage. Transcriptomic signatures showed that the signature of MA-5 shows an inverse correlation with aging and mortality and is oriented in the same direction as the OSKM-related iPSCs, suggesting the modification of aging pathways. Oral administration of MA-5 to mitochondrial disease model mouse showed increased survival. Our findings suggest that, in addition to enhancing ATP levels, the coordinated regulation of NAD+ metabolism, SIRT protein expression, and DNA-PK activity-constituting a novel therapeutic triad may contribute to the amelioration of hearing impairment and mitochondrial dysfunction, thereby improving life prognosis.

cell biology↗

β-Arrestin Condensates Regulate G Protein-Coupled Receptor Function

G protein-coupled receptors (GPCRs) are the largest class of receptors in the genome and control many signaling cascades essential for survival. GPCR signaling is regulated by {beta}-arrestins, multifunctional adapter proteins that direct receptor desensitization, internalization, and signaling. While at many GPCRs, {beta}-arrestins interact with a wide array of signaling effectors, it is unclear how {beta}-arrestins promote such varied functions. Here we show that {beta}-arrestins undergo liquid-liquid phase separation (LLPS) to form condensates that regulate GPCR function. We demonstrate that {beta}-arrestin oligomerization occurs in proximity to the GPCR and regulates GPCR functions such as internalization and signaling. This model is supported by a cryoEM structure of the adhesion receptor ADGRE1 in a 2:2 complex with {beta}-arrestin 1, with a {beta}-arrestin orientation that can promote oligomerization. Our work provides a paradigm for {beta}-arrestin condensates as regulators of GPCR function, with LLPS serving as an important promoter of signaling compartmentalization at GPCRs.

cell biology↗

Immune response to co-administration of bovine tuberculosis and contraceptive vaccines in badgers (Meles meles)

Controlling bovine tuberculosis (bTB) in livestock is often hindered by the presence of a wildlife reservoir, such as the Eurasian badger in the UK and Ireland. Vaccinating badgers against bTB can reduce the severity of Mycobacterium bovis infection and potential onwards transmission to cattle, badgers and other species, thus combined with population control might provide additional benefits (and reference models). To evaluate the effects of co-administration of a bTB vaccine (BCG) and a contraceptive vaccine (GonaCon), captive badgers were injected intramuscularly with either BCG only, GonaCon only or BCG+GonaCon (phase 1). The duration of immune response to BCG vaccination and booster vaccination was also evaluated in the BCG only group over a 24-month period (phase 2). The cellular immune responses to purified protein derivatives PPDA and PPDB, used to measure the effects of BCG, did not differ significantly between the BCG only and BCG+GonaCon badgers but were significantly different between the BCG only and GonaCon only badgers. Anti-GnRH antibody titres, quantified to measure the effect of GonaCon, peaked at the first sampling point for the majority of badgers in the GonaCon only and BCG+GonaCon groups. Throughout phase 1, lasting three months post-vaccination, badgers in both GonaCon treatment groups maintained high anti-GnRH antibody titres (1:128,000 or above) that are associated with infertility in this species, with no significant difference between the two GonaCon groups. The immune responses to BCG vaccination in the BCG only badgers (phase 2) were elevated at the first sampling point, approximately three weeks after initial vaccination and booster vaccination, generally declining thereafter. The results from the current study suggest that co-administration of BCG and GonaCon do not have notable deleterious effects on either of the vaccines and that the protective effect of BCG is enhanced by booster vaccination. The protective effects afforded by both vaccines when given together, in terms of reduced disease burden or fecundity, should be determined by further studies.

immunology↗

Chronic stress antagonizes formation of Stress Granules

Chronic stress mediates cellular changes that can contribute to human disease. However, fluctuations in RNA metabolism caused by chronic stress have been largely neglected in the field. Stress granules (SGs) are cytoplasmic ribonucleoprotein condensates formed in response to stress-induced inhibition of mRNA translation and polysome disassembly. Despite the broad interest in SG assembly and disassembly in response to acute stress, SG assembly in response to chronic stress has not been extensively investigated. In this study, we show that cells pre-conditioned with low dose chronic (24-hour exposure) stresses such as oxidative stress, endoplasmic reticulum stress, mitochondrial stress, and starvation, fail to assemble SGs in response to acute stress. While translation is drastically decreased by acute stress in pre-conditioned cells, polysome profiling analysis reveals the partial preservation of polysomes resistant to puromycin-induced disassembly. We showed that chronic stress slows down the rate of mRNA translation at the elongation phase and triggers phosphorylation of translation elongation factor eEF2. Polysome profiling followed by RNase treatment confirmed that chronic stress induces ribosome stalling. Chronic stress-induced ribosome stalling is distinct from ribosome collisions that are known to trigger a specific stress response pathway. In summary, chronic stress triggers ribosome stalling, which blocks polysome disassembly and SG formation by subsequent acute stress. Significant statementsStress granules (SGs) are dynamic cytoplasmic biocondensates assembled in response to stress-induced inhibition of mRNA translation and polysome disassembly. SGs have been proposed to contribute to the survival of cells exposed to toxic conditions. Although the mechanisms of SG assembly and disassembly in the acute stress response are well understood, the role of SGs in modulating the response to chronic stress is unclear. Here, we show that human cells pre-conditioned with chronic stress fail to assemble SGs in response to acute stress despite inhibition of mRNA translation. Mechanistically, chronic stress induces ribosome stalling, which prevents polysome disassembly and subsequent SG formation. This finding suggests that chronically stressed or diseased human cells may have a dysfunctional SG response that could inhibit cell survival and promote disease.

cell biology↗

Diet breadth in two polyphagous Spodoptera moths in a wide range of host and non-host plants and the potential for range expansion

O_LISeveral polyphagous moths are severe crop pests. Diet breadth patterns and mechanisms among polyphagous insects provide an excellent system to study ecological and evolutionary processes in herbivores, driving dietary specialization. However, studies of diet breadth on more than a handful of crops are scarce. C_LIO_LIHere, we estimated the diet breadth in two species of lepidopteran herbivores from the genus Spodoptera: S. littoralis (SL), with host range including both mono- and dicotyledonous plants and S. frugiperda (SF) Corn strain, primarily adapted to different grass species. C_LIO_LILarval performance on 23 crop and wild plant species from 17 families from terrestrial and wetland habitats was compared to an artificial diet in no-choice feeding bioassays. SL survived and performed better on most tested plants, particularly on the family level, except on two monocot plants (maize and leek), where SF performed well. There were five wild non-host plants where both generalists failed to survive. Nutrition indices assay corroborates the findings on a subset of plants. C_LIO_LIIn a subset of plants, larval feeding preference correlated partly, and larval attraction correlated well with larval performance. Female oviposition choice showed a weak correlation with larval performance. This weak correlation implies that these traits are decoupled, and other factors are crucial for female host plant selection. C_LIO_LIDuring larval dispersal greenhouse experiments, SL and SF larvae strongly tended to migrate onto their suitable host plants, indicating that this is one factor that modulates female host plant selection. C_LIO_LIIn summary, SL has a broader diet breath compared to SF, surviving on wild plants with no previous exposure. The present study provides the first comprehensive data on the diet breadth of two range-expanding and highly invasive polyphagous herbivores. C_LI

ecology↗

Study Design and Interim Analysis of the Cancer Lifetime Assessment Screening Study in Canines (CLASSiC): The First Prospective Cancer Screening Study in Dogs Using Next-Generation Sequencing-Based Liquid Biopsy

ObjectiveThe Cancer Lifetime Assessment Screening Study in Canines (CLASSiC) is a prospective, longitudinal cancer screening study, in which enrolled dogs are screened for cancer with physical exams and next-generation sequencing-based liquid biopsy testing on a serial basis. The goals of the first interim analysis, presented here, are to assess the benefits of using the OncoK9(R) liquid biopsy test as a cancer screening tool in a prospective clinical setting, and to demonstrate test performance for cancer detection, including preclinical detection. Subjects726 presumably cancer-free client-owned dogs were prospectively enrolled in the study across 24 clinical sites in the US and Canada. Most subjects were at high risk of cancer at the time of enrollment based on age and/or breed. 419 dogs that were enrolled for at least one year and had at least two cancer screening study visits, or that had received a definitive or presumptive diagnosis of cancer up to the time of the interim analysis, were included in the analysis. MethodsClinical data and a blood sample were collected at each study visit (once or twice per year and when cancer was clinically suspected). Cell-free DNA extracted from plasma was tested by OncoK9(R) using next-generation sequencing (NGS) technology. Results417 dogs were eligible for inclusion in the interim analysis and had classifiable outcomes, with a mean on-study duration of 422 days. Of these, 51 dogs were newly diagnosed with cancer (37 definitive, 14 presumptive), translating to a 12% (51/417) observed incidence over the study period; the liver, skin, bone, heart, spleen, lung, and lymph node(s) were the most common anatomic locations for disease. The prospectively observed sensitivity (detection rate) of the test was 56.9% (95% CI: 42.3-70.4%) with a specificity of 98.9% (95% CI: 97.0-99.6%). The prospectively observed positive predictive value was 87.9% (95% CI: 70.9-96.0%) and the negative predictive value was 94.3% (95% CI: 91.3-96.3%). NGS-based liquid biopsy doubled the overall number of cancer cases detected in this study population (from 25 to 51); remarkably, the detection rate for preclinical cancer was increased 4.6-fold from 12% (6/51) by routine care alone to 55% (28/51) by combining routine care with OncoK9(R) testing. Clinical RelevanceCLASSiC is the [fi]rst study to prospectively document the incidence of cancer in a predominantly high-risk canine population, and to prospectively demonstrate that the addition of NGS-based cancer screening to regularly scheduled wellness visits has the potential to substantially increase preclinical cancer detection in this population.

genomics↗

refMLST: Reference-based Multilocus Sequence Typing Enables Universal Bacterial Typing

SummaryCommonly used approaches for genomic investigation of bacterial outbreaks, including SNP and gene-by-gene approaches, are limited by the requirement for curated allele schemes. As a result, they only work on a select subset of known organisms, and fail on novel or less studied pathogens. We introduce refMLST, a gene-by-gene approach using the reference genome of a bacterium to form a scalable, reproducible and robust method to perform outbreak investigation. When applied to 1263 Salmonella enterica genomes, refMLST enabled consistent clustering, improved resolution and faster processing in comparison to chewieSnake. refMLST is applicable to any bacterial species with a public genome, does not require a curated scheme, and automatically accounts for genetic recombination. Availability and ImplementationrefMLST is freely available for academic use at https://bugseq.com/academic.

bioinformatics↗

Stress promotes RNA G-quadruplex folding in human cells

Guanine-rich nucleic acids can fold into G-quadruplex (G4) structures. Although endogenous RNAs contain sequences that can fold into RNA G4s (rG4s) in vitro, their folding and functions in vivo are not well understood. We show that the folding of putative rG4s in human cells into bona fide rG4 structures is dynamically regulated by stress. By using a high-throughput approach based on differential reactivity of dimethyl sulfate (DMS) towards Gs within folded vs unfolded rG4s, we identified hundreds of endogenous rG4s whose folding is promoted by cellular stress and validated them using a newly developed rG4-specific ligand. Stress-dependent rG4s are enriched in mRNA 3'-untranslated regions, suggesting their role in regulating mRNA stability under stress. Lastly, rG4 folding is reversible upon stress removal or adaptation. Our study show that rG4s function as regulatory elements in regulating mRNA stability and cellular stress response. One-Sentence SummaryRNA G-quadruplexes assemble under stress in human cells

molecular biology↗

What should a poor mother do? Influence of host plant quality on oviposition strategy and behavior in a polyphagous moth

To maximise fitness, individuals may apply different reproductive strategies. Such strategies could be phenotypically plastic and vary depending on the environment. For example, when resources are limited females often face a trade-off between investing in offspring quantity and quality, and how she balances this trade-off may depend on the environment. For phytophagous insects, and especially generalist insects, variation in host plant quality could have large effects on mating, reproduction and offspring performance. Here, we study if the polyphagous moth Spodoptera littoralis, which selects host plants through experience-based preference induction, also has a flexible allocation between egg weight and egg number as well as in temporal egg-laying behavior depending on larval host plant species. We found that S. littoralis has a canalized egg size and that an increased reproductive investment is made in egg quantity rather than egg quality. This increased investment depends on larval host plant species, probably reflecting parental condition. The constant egg weight may be due to physiological limitations or to limited possibilities to increase fitness through larger offspring size. We furthermore found that differences in onset of egg-laying is mainly due to differences in mating propensity between individuals raised on different host plant species. Thus, females do not seem to make a strategic reproductive investment in challenging environments. Instead, the low-quality host plant induces less and later reproduction, which could have consequences for population dynamics in the field.

ecology↗

Transcriptomic Insights into the Responses in Leaves Storing Lipid Organelles under Different Irradiances

To increase the nutritional value of forage, transgenic ryegrass known as High Metabolizable Energy (HME) were previously generated that co-express cysteine-oleosin and diacylglycerol O-acyltransferase. HME not only accumulate lipids in the leaf but also has elevated CO2 assimilation and increased biomass. Shading is one of the most influencing factors for ryegrass growth environments particularly in swards. The aim of this study, therefore, was to determine the influence of irradiance levels on photosynthesis and gene expression in the HME leaves when compared with their corresponding non-transformant (NT). Under low light (150-250 {micro}mol m-2 s-1) and standard light (600-1000 {micro}mol m-2 s-1), the HME accumulated more lipid than NT. The previously reported elevated photosynthesis and increased biomass was observed when the HME were grown under standard light but not under low light. Under both light conditions, compared to NT, the HME had upregulated a number of transcripts involved in lipid metabolism, light capturing, photosynthesis, and sugar signalling network while downregulated genes participated in sugar and fructan biosynthesis. We further discuss how the HME differentially manipulated several genes other metabolic pathways including maintenance of redox homeostasis. Combined, the data suggests that the increased photosynthesis capacity in the HME likely corresponds to an increase of micro-lipid sink strength; these are influenced by available light energy and may be related to diffusional and biochemical activities of stomata. Overall, this work provides a clearly understanding of the changes in molecular and biochemical mechanisms underlying the carbon storing as leaf lipid sink of the HME ryegrass. One sentence summaryShading led to increase leaf lipid accumulation but limit the greater photosynthesis trait of high lipid ryegrass which may be related to limitation of biochemical activities of stomata.

molecular biology↗

Caprin-1 binding to the critical stress granule protein G3BP1 is regulated by pH

G3BP is the central hub within the protein-RNA interaction network of stress-induced bio-molecular condensates known as stress granules (SG). The SG-associated proteins Caprin-1 and USP10 exhibit mutually exclusive binding to the structured NTF2-domain of G3BP1, thereby regulating G3BP1-mediated condensation, but with opposite effects: Caprin-1 promotes but USP10 inhibits SG formation. Herein, we present the crystal structure of G3BP1-NTF2 in complex with a Caprin-1 derived short linear motif (SLiM), which provides a molecular understanding for the mutually exclusive binding of USP10 and Caprin-1 to G3BP1. Caprin-1 but not USP10 contacts two G3BP1-NTF2 histidine residues, which was confirmed using biochemical, biophysical and cellular biological binding assays. G3BP1/Caprin-1 interactions disrupted via point mutations resulted in fewer and smaller SG condensates. In addition, biochemical binding assays demonstrated reduced binding of Caprin-1 to G3BP1 at lower pH values. Finally, ratiometric pH sensitive measurements of SGs revealed a substantial drop in pH compared to the adjacent cytosol, suggesting that reduced pH can fine-tune and regulate the G3BP1-mediated interaction network via a NTF2-mediated pH-sensitive SLiM-selection mechanism.

biochemistry↗

A Novel Stress Response Pathway Regulates Ribosome Biogenesis

Production of ribosomes is an energy-intensive process owing to the intricacy of these massive macromolecular machines. Each human ribosome contains 80 ribosomal proteins and four non-coding RNAs. Accurate assembly requires precise regulation of protein and RNA subunits. In response to stress, the integrated stress response (ISR) rapidly inhibits global translation. How rRNA is coordinately regulated with the rapid inhibition of ribosomal protein synthesis is not known. Here we show that stress specifically inhibits the first step of rRNA processing. Unprocessed rRNA is stored within the nucleolus, and, when stress resolves, it re-enters the ribosome biogenesis pathway. Retention of unprocessed rRNA within the nucleolus aids in the maintenance of this organelle. This response is independent of the ISR or inhibition of cellular translation but represents an independent stress-response pathway that we term Ribosome Biogenesis Stress Response (RiBiSR). Failure to coordinately regulate ribosomal protein translation and rRNA production results in nucleolar fragmentation. Our study unveils a novel stress response pathway that aims at conserving energy, preserving the nucleolus, and prevents further stress by regulation of rRNA processing.

biochemistry↗

Identifying states of collateral sensitivity during the evolution of therapeutic resistance in Ewings sarcoma

Advances in the treatment of Ewings sarcoma (EWS) are desperately needed, particularly in the case of metastatic disease. A deeper understanding of collateral sensitivity, where the evolution of therapeutic resistance to one drug aligns with sensitivity to another drug, may improve our ability to effectively target this disease. For the first time in a solid tumor, we produced a temporal collateral sensitivity map that demonstrates the evolution of collateral sensitivity and resistance in EWS. We found that the evolution of collateral resistance was predictable with some drugs, but had significant variation in response to other drugs. Using this map of temporal collateral sensitivity in EWS, we can see that the path towards collateral sensitivity is not always repeatable, nor is there always a clear trajectory towards resistance or sensitivity. Identifying transcriptomic changes that accompany these states of transient collateral sensitivity could improve treatment planning for EWS patients.

evolutionary biology↗

Spatio-temporal Proteomic Analysis of Stress Granule disassembly using APEX Reveals Regulation by SUMOylation and links to ALS pathogenesis

Cytoplasmic stress granules (SG) form in response to a variety of cellular stresses by phase-separation of proteins associated with non-translating mRNAs. SG provide insight into the biology of neurodegeneration, including amyotrophic lateral sclerosis (ALS) because they approximate some of the molecular conditions for nucleation of insoluble aggregates in neuropathological inclusions. Whereas much has been learned about SG formation, a major gap remains in understanding the compositional changes SG undergo during normal disassembly and under disease conditions. Here, we address this gap by proteomic dissection of SG temporal disassembly sequence, using multi-bait APEX proximity-proteomics. We discover 109 novel SG-proteins and characterize at proteomic resolution two biophysically distinct SG substructures. We further demonstrate that dozens of additional proteins are recruited to SG specifically during disassembly, indicating that it is a highly regulated process. The involved proteins link SG disassembly, to mitochondrial biology and the cytoskeleton. Parallel analysis with C9ORF72-associated dipeptides, which are found in patients with ALS and frontotemporal dementia, demonstrated compositional changes in SG during the course of disassembly and focused our attention on the roles SUMOylation in SG disassembly. We demonstrate that broad SUMOylation of SG-proteins is required for SG disassembly and is impaired by C9ORF72-associated dipeptides, representing an unexplored potential molecular mechanism of neurodegeneration. Altogether, out study fundamentally increases the knowledge about SG composition in human cells by dissecting the SG spatio-temporal proteomic landscape, provides an in-depth resource for future work on SG function and reveals basic and disease-relevant mechanisms of SG disassembly. HighlightsO_LIMulti bait APEX proximity labelling reveals 109 novel SG proteins and two distinct SG substructures. C_LIO_LIProteomic dissection of SG temporal disassembly under basal conditions and with a model of neurodegeneration. C_LIO_LIDisassembly-engaged proteins (DEPs) include SUMO ligases that are recruited during normal disassembly and dysregulated in ALS-like conditions. C_LIO_LIPervasive SG protein SUMOylation during SG disassembly is impaired by ALS-like conditions. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=168 SRC="FIGDIR/small/830133v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@15899daorg.highwire.dtl.DTLVardef@1dcd399org.highwire.dtl.DTLVardef@c7ef12org.highwire.dtl.DTLVardef@20c096_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

Regulation of FXR1 by alternative splicing is required for muscle development and controls liquid-like condensates in muscle cells

Fragile-X mental retardation autosomal homolog-1 (FXR1) is a muscle-enriched RNA-binding protein. FXR1 depletion is perinatally lethal in mice, Xenopus, and zebrafish; however, the mechanisms driving these phenotypes remain unclear. The FXR1 gene undergoes alternative splicing, producing multiple protein isoforms and mis-splicing has been implicated in disease. Furthermore, mutations that cause frameshifts in muscle-specific isoforms result in congenital multi-minicore myopathy. We observed that FXR1 alternative splicing is pronounced in the serine and arginine-rich intrinsically-disordered domain; these domains are known to promote biomolecular condensation. Here, we show that tissue-specific splicing of fxr1 is required for Xenopus development and alters the disordered domain of FXR1. FXR1 isoforms vary in the formation of RNA-dependent biomolecular condensates in cells and in vitro. This work shows that regulation of tissue-specific splicing can influence FXR1 condensates in muscle development and how mis-splicing promotes disease.\n\nHIGHLIGHTSO_LIThe muscle-specific exon 15 impacts FXR1 functions\nC_LIO_LIAlternative splicing of FXR1 is tissue- and developmental stage specific\nC_LIO_LIFXR1 forms RNA-dependent condensates\nC_LIO_LISplicing regulation changes FXR1 condensate properties\nC_LI

cell biology↗