CRISPR-based genetic screens in human pluripotent stem cells derived neurons and brain organoids

Cell and Tissue Research, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 25, 2024

Language: Английский

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hnRNPH1 Inhibits Influenza Virus Replication by Binding Viral RNA

Microorganisms, Journal Year: 2024, Volume and Issue: 13(1), P. 24 - 24

Published: Dec. 26, 2024

During the life cycle of influenza virus, viral RNPs (vRNPs) are transported to nucleus for replication. Given that a large number progeny RNA occupies nucleus, whether there is any host protein located in recognizes and inhibits replication remains largely unknown. In this study, explore role hnRNPH1 virus infection, we knocked down over-expressed proteins 293T cells, then infected cells with virus. The results showed H1N1 H9N2 viruses by restraining polymerase activity viruses. contains two recognition motifs (RRM1) RRM2. Further studies indicated specifically binds PB1, PA, NP genes. Mutation key residues tryptophan tyrosine RRM1 RRM2 abolished binding affinity suppression All suggested suppresses RNA.

Language: Английский

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Stabilizing mammalian RNA thermometer confers neuroprotection in subarachnoid hemorrhage

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 17, 2024

Summary Mammals tightly regulate their core body temperature, yet how cells sense and respond to small temperature changes at the molecular level remains incompletely understood. Here, we discover a significant enrichment of RNA G-quadruplex (rG4) motifs around splice sites cold-repressed exons. These thermosensing structures, when stabilized, mask sites, reducing exon inclusion. Focusing on cold-induced neuroprotective RBM3, demonstrate that rG4s near poison are stabilized low temperatures, leading exclusion. This enables evasion nonsense-mediated decay, increasing RBM3 expression cold. Additionally, intracellular potassium concentration stabilizes enhances expression, RBM3-dependent neuroprotection in mouse model subarachnoid hemorrhage. Our findings unveil mechanism mammalian RNAs directly perturbations, integrating them into gene programs. opens new avenues for treating diseases arising from splicing defects disorders benefiting therapeutic hypothermia. Highlights enriched cassette exons repressed upon cold shock act as thermometers mammals by controlling accessibility rG4 stability mediates temperature-dependent ex vivo Stabilizing 3a reduces brain damage hemorrhage Graphical abstract Here is refined G-quadruplexes acting evolutionarily conserved thermo- sensors, modulating alternative mammals. (rG4s) function reversible impacting dynamics. In temperatures or high conditions, can surrounding rendering these inaccessible, thereby promoting skipping. Conversely, under become destabilized, allowing be exposed, facilitating efficient well-known protein with functions, harbors that, inclusion, triggers NMD (non-sense mediated decay) mRNA. Under shield skipping increased expression. Stabilization through K + promotes skipping, enabling escape NMD, ultimately elevating Notably, 4-AP, clinically used pan voltage-gated channel blocker, protects against neuronal an RBM-dependent manner. (ISS: intronic silencer, ESE: enhancer, ACA: anterior cerebral artery, MCA: middle PPA: pterygopalatine ICA: internal carotid ECA: external CCA: common artery)

Language: Английский

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The cold-inducible RNA-binding protein RBM3 stabilises viral mRNA at the cooler temperatures of the upper respiratory tract

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: July 25, 2024

Abstract Temperature regulation is a key aspect of homeostasis, and several systems are involved in orchestrating adjustments gene expression at the cellular level. One such factor RBM3, cold-inducible RNA-binding protein implicated aspects mRNA processing regulation. The upper respiratory tract serves as unique environment regarding temperature Physiologically, lower relatively stable 37°C, while fluctuates or below 33°C. Adapting to this differential, subsequent differences transcriptome proteome, essential for viruses that infect cause disease simultaneously replicating transmitting from tract. At present, our understanding molecular mechanisms underlying influenza virus infection cooler temperatures lacking. Unsurprisingly, RBM3 levels highest nasopharyngeal tissue. Coupled with its known role positively regulating bound RNA, it an appealing candidate manipulation by viruses. We found siRNA knockdown significantly decreased viral replication. To disentangle direct effect RMB3 shift global colder temperatures, we generated A549 cell line constitutively overexpressing 37°C. Overexpression resulted significant increase was readily bind NP during prolong half-life these transcripts. In contrast, RNA binding null mutant reverses phenotype, validating interaction has stabilising on proviral nature increased further validated more clinically relevant model well-differentiated primary nasal epithelial cells. These data suggest supporting replication Understanding IAV could prove fundamental elucidating transmission reassortment. Author Summary establish productive infection, must overcome adapt within body. obstacle gradient tract, which route transmission, provides drastically different compared due growth temperatures. Here, detail investigation into (RBP) landscape between 33°C Our aim identify specific RBPs upregulated explore their A (IAV) lifecycle, thereby advancing knowledge Through combination virology mass spectrometry, identified RBP, important post-transcriptional regulator nucleoprotein (NP) mRNA. show binds specifically mRNA, ultimately promoting production infectious virions, abolishing capabilities reversed effect. Overall, find enhanced seen influences

Language: Английский

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Thermoregulated transcriptomics: the molecular basis and biological significance of temperature-dependent alternative splicing

Biochemical Journal, Journal Year: 2024, Volume and Issue: 481(15), P. 999 - 1013

Published: July 31, 2024

Temperature-dependent alternative splicing (AS) is a crucial mechanism for organisms to adapt varying environmental temperatures. In mammals, even slight fluctuations in body temperature are sufficient drive significant AS changes concerted manner. This dynamic regulation allows finely tune gene expression and protein isoform diversity response cues, ensuring proper cellular function physiological adaptation. Understanding the molecular mechanisms underlying temperature-dependent thus provides valuable insights into intricate interplay between stimuli regulation. this review, we provide an overview of recent advances understanding temperature-regulated across various biological processes systems. We will discuss machinery sensing translating cues changed patterns, adaptation regulatory extreme temperatures, role shaping transcriptome, functional implications development potential therapeutics targeting temperature-sensitive pathways.

Language: Английский

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Rbm3 deficiency leads to transcriptome-wide splicing alterations

RNA Biology, Journal Year: 2024, Volume and Issue: 21(1), P. 1 - 13

Published: Oct. 10, 2024

(RNA-binding motif protein 3) is a stress responsive gene, which maintains cellular homeostasis and promotes survival upon various harmful stimuli. Rbm3 shows conserved structural molecular similarities to heterogeneous nuclear ribonucleoproteins (hnRNPs), regulate all steps of the mRNA metabolism. Growing evidence pointing towards broader role in gene expression. Here, we demonstrate that deficiency linked transcriptome-wide pre-mRNA splicing alterations, can be reversed through co-expression from cDNA. Using an MS2 tethering assay, show regulates splice site selection similar other hnRNP proteins when recruited between two competing 5

Language: Английский

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