Search bioRxiv⌕ Search

Biology subjects

Roy, S.

Publications and source records attributed to Roy, S..

At least 19 recordsLinked to original sources

Discrete-time quantum walks reveal critical residues involved in long-range interactions in proteins

Abstract: Long-range interactions between residues in proteins underlie diverse biological processes such as signal transmission, conformational regulation, and catalysis. Identifying residues that mediate such communication remains a major challenge. Herein, we introduce a framework based on quantum walks and entropy, and employ it on protein structures to predict important residues participating in long-range interactions. The selected proteins have been well-studied experimentally in literature. Our approach models information flow through protein contact networks using discrete-time quantum walk dynamics, where quantum superposition and interference govern transition amplitudes at the residue level. Normalized time-averaged transition probabilities quantify interaction strength between residues, while Shannon entropy calculations identify residues exhibiting delocalized interaction behavior. We find that the residues identified by us are known to be structurally and functionally important in literature. We experimentally validate our formalism on AsLOV2 (PDB: 2V0U) using site-directed mutagenesis, protein purification, and dark-recovery kinetics. The predicted residues are important for understanding the mechanism of photosensing in AsLOV2. Significance: This study bridges quantum transport theory and structural biology by introducing discrete-time quantum walks as a mechanism to model communication between residues. We provide a scalable and interpretable method to predict key residues, without requiring molecular dynamics or prior functional annotation. The present method, inspired by quantum physics and quantum information, and validated by experiments, offers a broad and robust framework to study communication in biomolecular systems. It further demonstrates how quantum principles such as coherence and interference can enhance our understanding of information flow in biological macromolecules.

systems biology↗

Mechanistic insights into proteolytic activation ofmembrane-bound NAC transcription factor SlNACMTF3 intomato plants under heat stress

Stress is perceived at the cellular membrane, where membrane-bound transcription factors (MTFs) are activated, initiating a rapid gene-regulation cascade. We previously demonstrated the multifaceted role of tomato MTF, SlNACMTF3, in regulating expression of biotic and abiotic stress responsive genes. However, the molecular mechanism underlying its activation following stress perception remained unknown. Here, we show that the membrane rhomboid protease SlRP3 recognizes an ISK motif near the transmembrane domain of SlNACMTF3 for cleavage and releasing the transcription factor from the membrane. SlRP3 and SlNACMTF3 co-localize within the same plasma membrane microdomain and interact directly. Molecular dynamics simulations supported the proposed mechanism, in which heat-induced conformational switching of SlRP3, driven by reorientation of the L5 loop, enhances substrate accessibility and proteolytic activity. Furthermore, mutation of key SlRP3 residues involved in substrate interaction impaired SlNACMTF3 activation and reduced the expression of stress33 responsive genes. Analysis with SlNACMTF3-overexpressing tomato lines and SlRP3 transient knockdown leaves further revealed that transcription factor activation, stress35 responsive gene regulation, and cellular ROS detoxification occur coordinately. Collectively, these findings provide mechanistic insight into stress-triggered SlNACMTF3 activation and identify SlRP3 as a novel membrane-associated stress sensor that activates a major transcriptional regulator to enhance stress preparedness in economically important tomato plants.

plant biology↗

Human prion proteins and mutants through the lens of systems biology

Autocatalytic conformational changes of cellular prion proteins (PrPC) to the {beta}-sheet-enriched scrapie isoform (PrPSC) drive prion diseases, yet the precise molecular mechanisms remain unclear. To address this, we employ a multi-pronged framework integrating network-theoretic analyses, molecular dynamics (MD) simulations, and experimental validation to characterize critical residues governing human PrP (huPrP) structural integrity. Residue Interaction Networks (RINs) of: (A) wild-type (WT) huPrP, and, (B) five disease-associated point mutants (Met166Val, Glu220Lys, Glu200Lys, Asp178Asn, and Ser170Asn) were evaluated using three rigorous network methods: centrality measures, Shortest Path Alteration Fraction (SPAF), and Differential Networks (DN). Across all the diverse point mutations, five residues (Met206, Val209, Val210, Met213, and Cys214) consistently exhibit high centrality scores. SPAF and DN analyses demonstrate that alterations at these positions disrupt connectivity and increase the propensity for misfolding and protease resistance. To validate these topological findings, key residues within the C-terminal domain (121-230) were substituted via site-directed mutagenesis. Thermal denaturation monitored by circular dichroism (CD) spectroscopy reveals that the Met213Ala substitution significantly lowers the melting temperature, Tm, relative to the WT, whereas the Val210Ala variant unfolds with a Tm comparable to the WT. Furthermore, MD simulations of de novo AlphaFold-predicted mutant structures corroborate these findings. Mutants exhibit greater structural flexibility and deviation, with average root Mean Square Deviations rising from 0.18 nm (WT) to 0.32 nm (Cys214Ala), 0.29 nm (Met206Ala), and 0.25 nm (Val210Lys). This integration of theoretical and experimental techniques provides a robust framework for identifying critical determinants of protein stability.

systems biology↗

Inflammation is an essential trigger for POLE-driven ultramutated colorectal cancer

Pathogenic variants in the exonuclease (proofreading) domain of DNA polymerase {varepsilon} (POLE) are frequently detected in colorectal cancers (CRC) with high tumor mutational burden (TMB), yet their functional role in intestinal tumorigenesis remains unknown. Here, we investigated the impact of mutation in POLE on intestinal homeostasis using a conditional mouse model expressing the PoleP286R pathogenic driver in intestinal epithelial cells. PoleP286R mutant mice developed invasive adenocarcinomas in the small intestine with incomplete penetrance and extremely long latency but did not spontaneously develop colorectal tumors. However, induction of colitis with dextran sodium sulfate (DSS) resulted in a striking phenotypic shift to colorectal adenocarcinomas at high frequency. Molecular and histological analyses revealed hyperactivation of Wnt/{beta}-catenin, NF-{kappa}B, ERK and STAT3 pathways, accompanied by markedly higher proliferation of intestinal epithelial cells, in DSS-treated PoleP286R mutant mouse colons. Whole genome sequencing of the tumor demonstrated exceptionally high TMB (>100 mutations/megabase) and characteristic trinucleotide signatures, consistent with human POLE tumors. Recurrent mutations in Apc and Ctnnb1 underscored the central role of Wnt signaling in POLE-driven hypermutated CRC. In addition, intestinal organoids derived from PoleP286R mutant mice exhibited accelerated growth and increased expression of pro-proliferative genes. Together, these findings highlight POLE as a gatekeeper of intestinal genomic integrity and identify inflammation as the key driver that unleashes POLE-mutant CRC. Furthermore, this work establishes a mechanistically faithful preclinical platform for studying the molecular pathogenesis of CRC and advancing therapeutic drug discovery.

cancer biology↗

To mix or to de-mix?: Cation-π suppresses heterogeneity in local structure and dynamics of heterotypic Condensates

Heterotypic biomolecular condensates can exhibit pronounced spatial and dynamic heterogeneity because chemically distinct components compete between self- and cross-association. Here, we use coarse-grained simulations to investigate how cation-{pi} interactions modulate this competition in condensates formed by peptides with systematic variation of hydrophobicity-to-charge ratios in binary peptide pairs. By independently tuning sequence heterogeneity, peptide composition, and cation-{pi} interaction strength, we establish how an additional short-range interaction reshapes condensate stability, internal organization, and molecular dynamics. In the absence of cation-{pi} interactions, highly heterogeneous peptide pairs form distinct core-shell architectures, with charge- and hydrophobicity-rich sequences preferentially occupying different regions of the condensate. Introducing cation-{pi} interactions progressively enhances phase stability and promotes molecular mixing, with stronger interactions substantially increasing the critical temperature and condensate density. Energetic analysis reveals that cation-{pi} interactions partially compensate for the loss of electrostatic and hydrophobic stabilization, providing an additional and tuneable source of cohesive interactions. This enhanced cross-association also suppresses spatial and dynamic heterogeneity, yielding more homogeneous peptide distributions, slower translational diffusion, and more uniform chain reconfiguration within the dense phase. Collectively, our results demonstrate that cation-{pi} interactions provide a molecular mechanism for coupling condensate stability to spatial organization and internal dynamics, enabling sequence heterogeneity and composition to be translated into distinct condensate states.

biophysics↗

Single-Cell Omics to Uncover Cell-Type-Specific Gene Regulatory Networks Driving Mucilage Production in Specialized Roots

Aerial roots of certain maize (Zea mays) and sorghum (Sorghum bicolor) genotypes secrete a carbohydrate-rich mucilage that sustains nitrogen-fixing bacteria, and the underground roots of most plants secrete mucilage at their tips. In both settings, border cells released from the root cap produce this mucilage, yet the genes controlling their formation remain largely unknown. We sequenced the RNA of about 23,000 nuclei from mucilage-secreting roots of maize, sorghum, and Medicago truncatula, sampled before and after secretion. Each species yielded 10 to 15 clusters, which we annotated with published markers and with new markers that we validated by in situ hybridization in the two cereals and by promoter-reporter fusions in M. truncatula. We then inferred modular and cell type-specific gene regulatory networks. Transcriptional programs diverged across the three species except in the quiescent center and the meristem, and in the clusters containing border cells, where a shared set of regulators emerged. Silencing three of five candidate regulators reduced border cell number, while silencing a partially overlapping set of three regulators reduced root diameter in M. truncatula. Silencing the C2H2 zinc finger protein MtrunA17_Chr1g0148421 also reduced border cell viability. A maize insertion mutant of bzip89 released fewer border cells than the wild type. These networks offer entry points for tuning mucilage production in cereal and legume crops.

plant biology↗

A Novel PD-L1 Splice Isoform Modulates β Cell Communication in Response to Interferon Signaling

{beta} cell expression of the immune checkpoint ligand PD-L1 (encoded by CD274) limits autoimmune {beta} cell destruction in type 1 diabetes (T1D). {beta} cell display PD-L1 not only at the cell surface but also secreted on extracellular vesicles (EVs), which bind PD-1 and restrain CD8+ T cell activation. {beta} cell IFN signaling is an early driver of T1D pathogenesis, yet the mechanisms linking IFN signaling to the fate of PD-L1 protein remain undefined. Here, we identify an IFN- and coxsackievirus-inducible alternatively spliced isoform of PD-L1, PD-L1{Delta}3, lacking cassette exon 3 and generated in human {beta} cells and islets in response to IFN signaling or viral infection. PD-L1{Delta}3 transcripts are elevated in islets from donors with single autoantibody positivity (AAB+) and T1D. Unlike full-length PD-L1, which localizes to the plasma membrane, PD-L1{Delta}3 is retained intracellularly and loses the capacity to bind PD-1. Functionally, PD-L1{Delta}3 fails to suppress CD8+ and CD4+ T cell proliferation, activation, and cytotoxic cytokine release, and is not efficiently sorted into EVs. Convergently, a germline CD274 splice-site variant (c.682+1G>A) found in siblings with neonatal T1D yields a protein with the same intracellular retention, reduced {beta} cell PD-1 binding, and reduced {beta} cell and circulating EV PD-L1. Together these findings define PD-L1{Delta}3 as an IFN-induced splice variant that diverts PD-L1 away from its immunoregulatory, EV-competent form, revealing a post-transcriptional axis that shapes {beta} cell immune communication.

cell biology↗

Gasdermin E couples mitochondrial stress to STING-driven neuronal pyroptosis during Chandipura virus encephalitis

Neurotropic RNA viruses are major causes of fatal encephalitis worldwide, yet how infected neurons transition from antiviral defense to inflammatory cell death is not well characterized. Chandipura virus (CHPV), an emerging neurotropic rhabdovirus, causes acute, rapidly progressive encephalitis with high case fatality in children, but the mechanisms underlying its neuropathogenesis remain poorly defined. Here, we demonstrate that CHPV suppresses canonical RNA virus sensing early but subsequently switches to a mitochondria-driven innate immune program that culminates in inflammatory cell death. Early infection of neuronal cells with CHPV was marked by reduced levels of the mitochondrial antiviral adaptor protein, MAVS and attenuation of type I and III interferon responses. As infection progressed, mitochondrial dysfunction promoted accumulation of mtROS, mitochondrial accumulation of cleaved GSDME and cytosolic mtDNA release, triggering STING activation, which coincided with robust neuroinflammation and pyroptotic cell death. Pharmacological inhibition or genetic silencing of STING markedly attenuated inflammatory signaling, prevented pyroptotic membrane rupture and protected neurons from cell death without significantly affecting viral replication. In contrast, GSDME depletion reduced both viral replication and neuronal death. Notably, GSDME depletion markedly attenuated STING phosphorylation, while STING depletion also reduced GSDME activation, revealing functional coupling between these pathways during CHPV-induced neuronal injury. Collectively, our findings identify a mitochondria-GSDME-STING axis linking early immune evasion to neuroinflammation during CHPV infection, revealing a previously unrecognized mechanism of inflammatory neuronal death in viral encephalitis and highlighting STING as a potential therapeutic target in certain CNS viral infections.

immunology↗

Molecularly Distinct Innexin Gap Junction Channels and Undocked Hemichannels Regulate Glia Morphology and Function in Caenorhabditis elegans

Glial cells across species orchestrate nervous system development, maintenance, and function through ionic and metabolic crosstalk. These cells express multiple gap junction and hemichannel components. However, the mechanisms by which glia communicate utilizing these channel components to support nervous system architecture and function remain poorly understood. By studying GLR glial cells in Caenorhabditis elegans, we show that individual glial cells employ distinct innexin channel configurations to support specific functional roles, utilizing discrete downstream cellular mechanisms. We find that while innexins UNC-7and UNC-9 function as gap junction channels, the innexin INX-18 operates independently to form undocked hemichannels in discrete domains within the same cells. In this combinatorial configuration, UNC-7/UNC-9 gap junctions specifically regulate Synaptobrevin/SNB-1 localization in RME neurons, as previously reported. However, both channel types function non-redundantly to maintain glial morphology by regulating the Calpain/CLP-4 and CDK-5 pathway through cell-autonomous modulation of intracellular calcium levels. In contrast, INX-18 hemichannels in GLR glia regulate high-salt-induced paralysis behavior through distinct downstream cellular mechanisms, potentially in conjunction with tyramine signaling. Altogether, our findings demonstrate a broader repertoire of innexin channel utilization at the individual glial cell level, supporting specific aspects of nervous system maintenance and functioning.

neuroscience↗

Multiple trans-regulators shape enhancer-promoter hub organization at a multi-enhancer locus

Developmental genes are frequently regulated by multiple enhancers distributed across large cis-regulatory regions. How these enhancers communicate with their target promoter and how their interactions are shaped by distinct developmental transcriptional environments remain incompletely understood. Here, we investigate the chromatin organization of the Drosophila shavenbaby locus, a developmental gene controlled by seven distal enhancers. Tissue-specific UMI-4C revealed extensive enhancer-promoter and enhancer-enhancer interactions, including in cell populations where individual enhancers are inactive. Quantitative three-dimensional DNA-FISH revealed compact enhancer-promoter hubs enriched in shavenbaby-expressing cells, yet also present in non-expressing cells and prior to expression. Perturbation of shavenbaby regulators, transcription factors, and architectural proteins revealed that multiple factors contribute to hub organization. Their relative contributions differed between epidermal populations, indicating that similar hubs can be supported by different combinations of regulators. Perturbations that reduced hub organization were frequently associated with reduced shavenbaby-dependent trichome formation. Together, our results identify a robust, multi-factorial enhancer-promoter hub that is shaped by distinct regulatory inputs across developmental contexts.

developmental biology↗

Engineering Heterotypic Biomolecular Condensates with Synthetic Peptides for Controlled Spatial Organization and Liquid-like Nature

Sequence heterogeneity is a defining feature of cellular biomolecular condensates, yet how competing interaction motifs encode their thermodynamic stability, internal organization, and dynamics remains poorly understood. Here, we systematically tune the hydrophobicity mismatch between intrinsically disordered peptide pairs to establish sequence hydrophobicity as a programmable determinant of heterotypic condensate behaviour. We show that heterotypic condensates are thermodynamically more stable than homotypic ones having same average hydrophobicity through the cooperative interplay of short-range hydrophobic and long-range electrostatic interactions. Increasing hydrophobicity mismatch drives a composition-dependent transition from homogeneous condensates to core-shell architectures accompanied by pronounced spatial and dynamical heterogeneity, whereas reducing sequence disparity restores homogeneous organization and nearly uniform dynamics. Our results establish a direct molecular link between sequence chemistry, phase stability, condensate architecture, and transport dynamics, providing predictive design principles for engineering synthetic biomolecular condensates with programmable organization and material properties. TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=105 SRC="FIGDIR/small/743201v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@12e7fborg.highwire.dtl.DTLVardef@13c31a0org.highwire.dtl.DTLVardef@de3453org.highwire.dtl.DTLVardef@3d4600_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

Amaranthus viridis-derived phytopriming reprograms redox homeostasis and limits arsenic accumulation in rice

Arsenic (As) poses a major threat to rice productivity and food safety due to its high bioaccumulation potential and subsequent entry into the human food chain. In rice, As impairs seed germination, disrupts morpho-anatomical development, and induces oxidative stress. This study evaluates seed priming with an aqueous extract of the agricultural weed Amaranthus viridis (AvE) as a sustainable strategy to alleviate As-induced phytotoxicity. AvE priming significantly improved germination (71-75%) and morpho-physiological performance under As stress. It reduced oxidative stress markers, including H2O2 (21-38%), malondialdehyde (13-26%), and proline (18.9-44.7%), while increasing antioxidant metabolites, polyphenols and glutathione by up to 2.34-fold and 41%, respectively. Microscopy confirmed restoration of cellular integrity and anatomical organisation in primed seedlings. ICP-OES analysis showed that AvE priming reduced root As uptake by up to 39%, root-to-shoot translocation by up to 58%, and grain As accumulation by up to 95% compared with unprimed plants. qRT-PCR revealed modulation of genes involved in As homeostasis, indicating coordinated physiological and transcriptional responses. Importantly, improved agronomic performance further demonstrated the translational potential of this approach. This study provides the first evidence that A. viridis extract is a cost-effective, sustainable biostimulant for producing low-As rice in contaminated regions.

plant biology↗

Molecular Detection and Antibiogram Assay of Escherichia coli Isolated from Poultry Farm Environments

BackgroundEscherichia coli is a common environmental and zoonotic bacterium that serves as an important indicator of fecal contamination and antimicrobial resistance (AMR) in poultry production systems. The emergence of multidrug-resistant (MDR) E. coli in poultry farm environments poses a significant One Health threat to animal, human, and environmental health. ObjectiveThis study aimed to isolate, identify, determine the prevalence, assess the antibiotic susceptibility pattern, and molecularly confirm Escherichia coli isolated from poultry farm environments in Lalmonirhat district, Bangladesh. Materials and MethodsA cross-sectional study was conducted from January to June 2024 using 60 environmental samples comprising water (n = 20), soil (n = 20), and bird-dropping (n = 20) samples collected from poultry farms in five upazilas of Lalmonirhat district. Isolation and identification of E. coli were performed using standard cultural, morphological, and biochemical techniques. Antimicrobial susceptibility was determined by the Kirby-Bauer disc diffusion method following CLSI guidelines. Ten randomly selected isolates were confirmed by polymerase chain reaction (PCR) targeting the species-specific uidA gene. ResultsThe overall prevalence of E. coli was 50% (30/60). Source-wise prevalence was highest in bird-dropping samples (75%), followed by water (45%) and soil (30%). Area-wise prevalence ranged from 33.33% in Patgram to 66.67% in Hatibandha. The isolates exhibited the highest resistance to oxytetracycline (66.67%) and ciprofloxacin (60%), while the highest susceptibility was observed to erythromycin (63.33%), neomycin (56.67%), and amoxicillin (56.67%). All ten isolates subjected to PCR produced the expected 486 bp uidA gene amplicon, confirming their identity as E. coli. ConclusionThe findings demonstrate a considerable prevalence of antimicrobial-resistant E. coli in poultry farm environments of Lalmonirhat district. Strengthening farm biosecurity, improving hygiene and waste management, implementing antimicrobial stewardship, and maintaining continuous molecular surveillance are essential to minimize the dissemination of resistant E. coli within a One Health framework.

microbiology↗

ZNF768 regulates expression of E2F1 protein to drive G0/G1 transition and cell cycle progression.

Accurate and tightly coordinated cell cycle progression and cell proliferation are critical for development, growth and homeostasis of an organism. Recently, Zinc finger protein 768 (ZNF768) was identified as a transcription factor driving cellular proliferation, in both a p53-dependent and independent manner. ZNF768 interacts with and represses p53 functions to limit cell cycle delay. Independently, ZNF768 promotes the transcription of key regulators of the cell cycle machinery, although the mechanisms through which this occurs remain unknown. Here, we report that ZNF768 protein levels are tightly regulated during the cell cycle, and its depletion leads to cell cycle exit and induction of quiescence. We found that ZNF768 modulates the cell cycle, at least in part, by controlling expression of the major pro-proliferative transcription factor E2F1 independently of p53 activation. Consequently, depletion of ZNF768, which also represses expression of the key mitotic transcription factor and E2F1 target FOXM1, leads to numerous mitotic errors. Supporting these findings, cancer genomics analyses reveal that ZNF768 expression levels are positively associated with E2F1 and FOXM1 expression levels in human tumors, suggesting that cancer cells might use ZNF768 to override cell cycle arrest, sustain proliferation, and promote cancer progression. Altogether, our results reveal that ZNF768 modulates cell cycle entry and proliferation, at least in part by regulating E2F1 expression.

cell biology↗

Assessing the physiological S nitrosoproteome reveals nitric oxide mediated regulatory networks in rice

Nitric oxide metabolism-based protein post-translational modifications, such as reversible S- nitrosylation, have been at the pinnacle of plant redox research owing to their significant correlation with seed dormancy, interaction with other signaling molecules, plant development and metabolism, biotic and abiotic plant stress responses, immune defense responses against plant pathogens, and senescence. The rapid interconversion of reactive nitrogen species, the abrogation of nitric oxide homeostasis by exogenous supplementation of NO donors and scavengers, the lack of spatio-temporal specificity of NO signaling, and the limited bioavailability or assay sensitivity for detection often limit the effectiveness of identifying and characterizing S-nitrosothiols in plants. Hitherto unknown, we report the first experimental evidence of the total in vivo S-nitrosoproteome in Oryza sativa L. subsp. indica, comprising 134 PSNOs, enriched with 169 putative sites susceptible to S-nitrosylation, without any exogenous supplementation of NO donors. In the present study, mercuric salt- driven facile decomposition of S-nitrosoproteins in the presence of nitrone spin trap 5,5- dimethyl-1-pyrroline N-oxide, resulting in the synthesis of DMPO-nitrone adducts with PSNO-derived protein thiyl radicals in O. sativa, has been demonstrated as an efficient and novel strategy for characterizing the PSNOs using mass spectrometry analysis. The evidence of physiological levels of PSNOs was further re-examined in a bi-directional qualitative and quantitative approach involving the 2,3-diaminonaphthalene assay in tandem with fluorescence-based visualization and fluorometric quantification. In silico analyses, involving both functional enrichment and pathway prediction analyses, have furthermore revealed unique protein-protein interaction networks and signaling pathways among the S- nitrosoproteome candidates and their predictable physiological roles in O. sativa indica, awaiting further in vitro validation for their functional correlation in response to S- nitrosylation. In conclusion, the present study provides novel evidence of nitric oxide signaling in rice cultivars under physiological conditions, bringing new insights into the potential in vivo transnitrosylation of regulatory or active-site cysteine thiols.

biochemistry↗

On-demand, reversible blood-brain barrier opening via electrical activation of piezoelectric nanoparticles for targeted brain drug delivery

Delivering therapeutics to the brain remains one of the most persistent challenges in medicine, because the blood-brain barrier (BBB) excludes over 98% of small-molecule drugs and virtually all biologics from the central nervous system (CNS). We developed electrical BBB modulation (eBBB), an on-demand platform combining vascular-targeting poly-L-lactic acid nanoparticles with high-definition transcranial direct current stimulation to achieve spatially and temporally controlled BBB opening. eBBB produced localized, reversible increases in BBB permeability confined to the stimulated cortex, with the opening area tunable via electrode geometry. This transient window enhanced regional delivery of a small-molecule drug, full-length immunoglobulins, and adeno-associated viral vectors, which are cargo classes otherwise completely excluded by the intact BBB. Neurovascular unit architecture was preserved with no lasting histological damage. Integrating a biodegradable nanomaterial with a clinically evaluated stimulation technology, eBBB offers a programmable, minimally invasive strategy for regional CNS drug delivery across brain malignancies and neurological disorders. One sentence summaryElectrical activation of piezoelectric nanoparticles reversibly opens the blood-brain barrier for minimally invasive drug delivery to targeted cortical regions.

bioengineering↗

PCIF1 loss licenses antitumour immunity via cholesterol biosynthesis

Cholesterol biosynthesis is classically viewed as a tumour supportive metabolic programme, yet cholesterol-associated metabolic states can paradoxically coincide with improved responses to immune checkpoint blockade (ICB)1, highlighting an unresolved context-dependent relationship between cholesterol homeostasis and antitumour immunity. Here we identify PCIF1, a cap-specific RNA methyltransferase2, as a tumour-intrinsic brake on the inflammatory potential of cholesterol biosynthesis. PCIF1 depletion had limited effects on tumour cell proliferation in vitro, but suppressed tumour growth in immunocompetent, not immunodeficient, hosts and sensitized immune-refractory syngeneic tumours to ICB. Immune profiling by single-cell RNA sequencing, flow cytometry and multiplex immunofluorescence revealed increased CD8+ T cell infiltration in PCIF1-deficient tumours, and CD8+ T cell depletion restored tumour growth. Mechanistically, PCIF1 loss selectively enhanced translational engagement of SCAP, activated SREBP2-dependent cholesterol biosynthesis and promoted intracellular cholesterol accumulation. Unexpectedly, activation of this cholesterol programme induced a tumour cell inflammatory cytokine state enriched for T cell-recruiting chemokines, including CXCL10. Pharmacological perturbation of the SCAP-cholesterol axis attenuated this inflammatory programme, placing cholesterol sensing upstream of PCIF1 loss-induced tumour cell inflammatory activation. In ICB-treated patient cohorts, high PCIF1 expression in pre- and on-treatment samples was associated with poor clinical response, and across tumour types, PCIF1 expression inversely correlated with cytolytic immune signatures. These findings reveal PCIF1-mediated SCAP translational control as a tumour cell checkpoint that constrains the antitumour inflammatory activity of a canonical growth-associated cholesterol programme and shapes checkpoint therapy responsiveness.

cancer biology↗

Structural Topology-based Electrostatic Model (STEM) Reveals Ion-Coordination Exchange as a Driver of RNA Folding Dynamics

RNA is a highly charged polyelectrolyte whose folding into functional architectures depends on an ionic atmosphere that screens strong electrostatic repulsion along the phosphate backbone. Whereas monovalent ions primarily stabilize secondary structure, divalent magnesium (Mg2+) drives tertiary folding often via site-specific and adopting various dynamic coordination modes. Current RNA structure-prediction frameworks rely largely on static direct-contact information, overlooking ion-mediated interactions and the dynamic exchange between distinct coordination modes--particularly the dynamic exchange between direct (inner) and solvent-separated (outer-sphere) Mg2+-phosphate coordination that often controls RNAs conformational transition. Here, we introduce the Structural-based Electrostatic Model (STEM), a hybrid implicit-explicit framework that explicitly captures how the dynamic exchange between distinct ion-coordination modes dictates folding pathways. STEM combines explicit Mg2+ ions to resolve site-specific interactions with implicit K+ ions to describe counter-ion condensation mediated electrostatic screening through generalized Manning counter-ion condensation model, enabling computationally efficient exploration of RNA folding landscapes. The model accurately reproduces crystallographic ion-binding sites, experimental preferential ion-interaction coefficients, and Small-Angle X-ray Scattering (SAXS)-derived radii of gyration across diverse RNA systems. Applied to a 58-nt rRNA fragment, STEM reveals that folding from an intermediate to the native state is driven by a chelated Mg2+-mediated tertiary contact and captures the resulting coordination-dependent conformational breathing. By shifting the paradigm from static direct-contact descriptions to ion-mediated dynamic interactions, STEM provides a physically grounded framework for predicting dynamic ensembles of RNA structures, resolving their folding free-energy landscapes, and elucidating the mechanisms of RNA folding and function beyond native conformations across physiological salt conditions. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/734987v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@27da3borg.highwire.dtl.DTLVardef@689310org.highwire.dtl.DTLVardef@18f32f0org.highwire.dtl.DTLVardef@593184_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗