Is -Associated Hearing Loss (DFNB93) a Gene Therapy Target? Preclinical Progress and Patient Registry.

modulates presynaptic Ca1.3 Ca channel function in inner hair cells (IHCs) and is required for indefatigable synaptic sound encoding. Biallelic variants in are associated with non-syndromic hearing loss (DFNB93).

On-Target Photoassembly of Pyronin Dyes for Super-Resolution Microscopy.

Controlled photoactivation is an auspicious and emerging approach in super-resolution microscopy, offering virtually zero background signal from the marker prior to activation. Pyronins are well-established fluorophores, but due to their inherent intercalating tendency towards nucleic acids, their use has been mostly avoided in super-resolution microscopy. Here, we describe a new class of diaryl ether and diaryl silane molecules that upon photoactivation close into fluorescent (silicon-)pyronins and term them Pyronin Upon Light Irradiation (PULI).

Neddylation regulates the development and function of glutamatergic neurons.

Neuronal development and function are orchestrated by a plethora of regulatory mechanisms that control the abundance, localization, interactions, and function of proteins. A key role in this regard is assumed by post-translational protein modifications (PTMs). While some PTM types, such as phosphorylation or ubiquitination, have been explored comprehensively, PTMs involving ubiquitin-like modifiers (Ubls) have remained comparably enigmatic (Ubls).

Presynaptic ATP Decreases During Physiological-Like Activity in Neurons Tuned for High-Frequency Transmission.

Recent evidence indicates that the concentration of ATP remains stable during neuronal activity due to activity-dependent ATP production. However, the mechanisms of activity-dependent ATP production remain controversial. To stabilize the ATP concentration, feedforward mechanisms, which may rely on calcium or the sodium-potassium pump, do not require changes in the ATP and ADP concentrations.

Dominant-negative NARS1 R534* mutation causes wild-type subunit poisoning and heterodimer predominance in cells.

Aminoacyl-tRNA synthetases (aaRSs) catalyze the aminoacylation of tRNA with their cognate amino acids, an essential step in protein biosynthesis. While biallelic mutations in aaRSs often result in severe multi-organ dysfunction accompanied by developmental delays, monoallelic mutations typically cause milder, tissue-specific symptoms. However, a de novo monoallelic nonsense mutation (R534*) in the asparaginyl-tRNA synthetase (AsnRS)-resulting in a premature stop codon and 15-residue C-terminal truncation-has been identified in multiple families and is associated with severe neurodevelopmental symptoms.

Cotranslational protein folding through non-native structural intermediates.

Cotranslational protein folding follows a distinct pathway shaped by the vectorial emergence of the peptide and spatial constraints of the ribosome exit tunnel. Variations in translation rhythm can cause misfolding linked to disease; however, predicting cotranslational folding pathways remains challenging. Here, we computationally predict and experimentally validate a vectorial hierarchy of folding resolved at the atomistic level, where early intermediates are stabilized through non-native hydrophobic interactions before rearranging into the native-like fold.

A Novel α-Synuclein K58N Missense Variant in a Patient with Parkinson's Disease.

Background: Parkinson's disease (PD) is a complex multifactorial disorder with a genetic component in about 15% of cases. Multiplications and point mutations in SNCA gene, encoding α-synuclein (aSyn), are linked to rare familial forms of PD.

Objective: Our goal was to assess the clinical presentation and the biological effects of a novel K58N aSyn mutation identified in a patient with PD.

Multiple Mammalian Cytokines and Erythropoietin-Mimetic Peptides Protect Insect Neurons via Phylogenetically Conserved Cytokine Receptor-Like Factor 3 (CRLF3).

Erythropoietin (Epo) and its non-erythropoietic splice variant EV-3 have demonstrated potent neuroprotective effects across species, although the respective mechanisms are variable and incompletely understood. Unlike vertebrates, insects lack both Epo and the classical Epo receptor but express Cytokine Receptor-Like Factor 3 (CRLF3), a conserved type I receptor that serves as a neuroprotective receptor for Epo and EV-3 in insects and human iPSC-derived neuron-like cells. Insects, which express CRLF3 but lack all other group 1 type I cytokine receptors, represent a suitable model to study the function of CRLF3 in neuroprotection.

A nanobody specific to prefusion glycoprotein B neutralizes HSV-1 and HSV-2.

The nine human herpesviruses, including herpes simplex virus 1 and 2, human cytomegalovirus and Epstein-Barr virus, present a significant burden to global public health. Their envelopes contain at least ten different glycoproteins, which are necessary for host cell tropism, attachment and entry. The best conserved among them, glycoprotein B (gB), is essential as it performs membrane fusion by undergoing extensive rearrangements from a prefusion to postfusion conformation.

Identification and validation of a tear fluid-derived protein biomarker signature in patients with amyotrophic lateral sclerosis.

The diagnosis of Amyotrophic Lateral Sclerosis (ALS) remains challenging, particularly in early stages, where characteristic symptoms may be subtle and nonspecific. The development of disease-specific and clinically validated biomarkers is crucial to optimize diagnosis. Here, we explored tear fluid (TF) as a promising ALS biomarker source, given its accessibility, anatomical proximity to the brainstem as an important site of neurodegeneration, and proven discriminative power in other neurodegenerative diseases.

Selective ubiquitination of drug-like small molecules by the ubiquitin ligase HUWE1.

The ubiquitin system regulates eukaryotic physiology by modifying myriad substrate proteins. Substrate specificity and the assembly of ubiquitin signals are determined by ubiquitin ligases, some of which also modify non-protein biomolecules. Here we expand this substrate realm, revealing that the human ligase HUWE1 can target drug-like small molecules.

Combining Correlative Cryogenic Fluorescence and Electron Microscopy and Correlative Cryogenic Super-Resolution Fluorescence and X-Ray Tomography-Novel Complementary 3D Cryo-Microscopy Across Scales to Reveal Nanoparticle Internalization Into Cancer Cells.

Understanding the intracellular fate of nanoparticles (NPs) is essential for advancing nanomedicine, particularly in targeted drug delivery for cancer therapy. Here, we present a complementary cryogenic microscopy workflow across scales to investigate the uptake and subcellular localization of zirconyl-containing inorganic-organic hybrid nanoparticles (IOH-NPs) in murine breast cancer cells. Our approach integrates cryogenic fluorescence microscopy (cryo-FM), cryo-focused ion beam scanning electron microscopy (cryo-FIBSEM), and cryo-soft X-ray tomography (cryo-SXT), enabling molecular specificity, high-resolution imaging, and volumetric ultrastructural analysis in near-native cellular states.

Translational repression by 4E-T is crucial to maintain the prophase-I arrest in vertebrate oocytes.

Meiotic maturation of vertebrate oocytes occurs in the near-absence of transcription. Thus, female fertility relies on timely translational activation of maternal transcripts stockpiled in full-grown prophase-I-arrested oocytes. However, how expression of these mRNAs is suppressed to maintain the long-lasting prophase-I arrest remains mysterious.

SynapseNet: Deep Learning for Automatic Synapse Reconstruction.

Electron microscopy is an important technique for the study of synaptic morphology and its relation to synaptic function. The data analysis for this task requires the segmentation of the relevant synaptic structures, such as synaptic vesicles, active zones, mitochondria, presynaptic densities, synaptic ribbons, and synaptic compartments. Previous studies were predominantly based on manual segmentation, which is very time-consuming and prevented the systematic analysis of large datasets.

Developmental transformations of Purkinje cells tracked by DNA electrokinetic mobility.

Brain development begins with neurogenesis in progenitor zones and ends with expansive, intricately-patterned cellular diversity in the adult brain. We took advantage of bioelectric interactions between DNA and embryonic tissue to perform "stereo-tracking," a developmental targeting strategy that differentially labels cells at different depths within progenitor zones. This 3D labeling was achieved by delivery of plasmids with distinct electrokinetic mobilities in utero.

Integrative bioinformatics and deep learning to identify common genetic pathways in Crohn's disease and ischemic cardiomyopathy.

Crohn's disease (CD) and ischemic cardiomyopathy (ICM) share inflammatory characteristics, yet their common genetic underpinnings remain underexplored. Using an integrative bioinformatics approach, we analyzed GEO datasets (GSE3365 and GSE9128) to identify shared genetic pathways between CD and ICM. Through differential expression analysis, we identified 60 common differentially expressed genes (CDEGs).

Nanoparticle-assisted dynamic nuclear polarization in liquids.

Gold nanoparticles (AuNPs) protected with self-assembled organic monolayers represent versatile nanostructures with applications in catalysis, molecular sensing, and recognition. The ordered arrangement of ligands within the monolayer promotes specific orientations and proximities of functional groups, facilitating weak yet selective interactions with small molecules in solution. In the context of nuclear magnetic resonance (NMR), AuNPs can act as magnetization reservoirs, allowing selective transfer to interacting molecules and making them promising chemosensors.

Translocations can drive expression changes of multiple genes in regulons covering entire chromosome arms.

Chromosomal translocations have largely been implicated in tumor development. However, beyond the consequences of aberrant gene expression near the breakpoint, their effects remain underexplored. In this work, we characterize the interplay between translocations, chromatin organization and gene expression using mantle cell lymphoma (MCL) as a model.

IWS1 positions downstream DNA to globally stimulate Pol II elongation.

The protein IWS1 (Interacts with SPT6 1) is implicated in transcription-associated processes, but a direct role in RNA polymerase (Pol) II function is unknown. Here, we use multi-omics kinetic analysis after rapid depletion of IWS1 in human cells to show that loss of IWS1 results in a global decrease of RNA synthesis and a global reduction in Pol II elongation velocity. We then resolve the cryo-EM structure of the activated Pol II elongation complex with bound IWS1 and elongation factor ELOF1 and show that IWS1 acts as a scaffold and positions downstream DNA within the cleft of Pol II.

Direct mechanical communication of cellular to nuclear shapes in oocytes.

The mechanical properties of the cytoplasm and nucleoplasm are crucial for the correct and robust functioning of a cell and play a key role in understanding how mechanical signals are transferred to the nucleus. Here, we demonstrate remarkable shape mimicry between the cellular and nuclear shape of oocytes, following the externally applied deformation without direct contact between the cell cortex and the nucleus. This effect arises from a surprisingly soft and fluid-like nucleoplasm that barely resists external strain, while the viscoelastic cytoplasm drives shape transmission.

Current practices in the study of biomolecular condensates: a community comment.

The realization that the cell is abundantly compartmentalized into biomolecular condensates has opened new opportunities for understanding the physics and chemistry underlying many cellular processes, fundamentally changing the study of biology. The term biomolecular condensate refers to non-stoichiometric assemblies that are composed of multiple types of macromolecules in cells, occur through phase transitions, and can be investigated by using concepts from soft matter physics. As such, they are intimately related to aqueous two-phase systems and water-in-water emulsions.

Synergistic activity of RSL3 and Pyrimethamine to inhibit the proliferation of .

Malaria tropica, caused by (), remains a global health challenge with limited therapeutic options. In mammalian cells, the small-molecule compound RAS-selective lethal 3 (RSL3) induces ferroptosis via lipid peroxidation. In this study, we demonstrate that RSL3 synergizes with Pyrimethamine, an inhibitor of dihydrofolate reductase (DHFR), to suppress parasite proliferation in red blood cells (RBCs).

Chemical crosslinking extends and complements UV crosslinking in analysis of RNA/DNA nucleic acid-protein interaction sites by mass spectrometry.

Ultraviolet (UV) crosslinking with mass spectrometry (XL-MS) has been established for identifying RNA- and DNA-binding proteins along with their domains and amino acids involved. Here, we explore chemical XL-MS for RNA-protein, DNA-protein, and nucleotide-protein complexes in vitro and in vivo. We introduce a specialized nucleotide-protein-crosslink search engine, NuXL, for robust and fast identification of such crosslinks at amino acid resolution.

Structure Determination from Single-Molecule X-ray Scattering Images Using Stochastic Gradient Ascent.

Scattering experiments using ultrashort X-ray free electron laser pulses have opened a new path for structure determination of a wide variety of specimens, including nanocrystals and entire viruses, approaching atomistic spatial and femtoseconds time resolution. However, random and unknown sample orientations as well as low signal-to-noise ratios have so far prevented a successful application to smaller specimens like single biomolecules. We here present resolution-annealed stochastic gradient ascent (RASTA), a new approach for direct atomistic electron density determination, which utilizes our recently developed rigorous Bayesian treatment of single-particle X-ray scattering.