bluuebunny/crossref_metadata_2025_split huggingface dataset:
m6A mRNA Methylation in Brown Adipose Tissue Regulates Systemic Insulin Sensitivity via an Inter-Organ Prostaglandin Signaling Axis
Authors: Xiao Ling , De Jesus Dario F. , Ju Cheng-Wei , Wei Jiang-Bo , Hu Jiang , DiStefano-Forti Ava , Tsuji Tadataka , Cero Cheryl , Männistö Ville , Manninen Suvi M. , Wei Siying , Ijaduola Oluwaseun , Blüher Matthias , Cypess Aaron M. , Pihlajamäki Jussi , Tseng Yu-Hua , He Chuan , Kulkarni Rohit N. 
Published: May 2023

SummaryBrown adipose tissue (BAT) has the capacity to regulate systemic metabolism through the secretion of signaling lipids. N6-methyladenosine (m6A) is the most prevalent and abundant post-transcriptional mRNA modification and has been reported to regulate BAT adipogenesis and energy expenditure. In this study, we demonstrate that the absence of m6A methyltransferase-like 14 (METTL14), modifies the BAT secretome to initiate inter-organ communication to improve systemic insulin sensitivity. Importantly, these phenotypes are independent of UCP1-mediated energy expenditure and thermogenesis. Using lipidomics, we identified prostaglandin E2 (PGE2) and prostaglandin F2a (PGF2a) as M14KO-BAT-secreted insulin sensitizers. Notably, circulatory PGE2 and PGF2a levels are inversely correlated with insulin sensitivity in humans. Furthermore,in vivoadministration of PGE2 and PGF2a in high-fat diet-induced insulin-resistant obese mice recapitulates the phenotypes of METTL14 deficient animals. PGE2 or PGF2a improves insulin signaling by suppressing the expression of specific AKT phosphatases. Mechanistically, METTL14-mediated m6A installation promotes decay of transcripts encoding prostaglandin synthases and their regulators in human and mouse brown adipocytes in a YTHDF2/3-dependent manner. Taken together, these findings reveal a novel biological mechanism through which m6A-dependent regulation of BAT secretome regulates systemic insulin sensitivity in mice and humans.HighlightsMettl14KO-BAT improves systemic insulin sensitivity via inter-organ communication;PGE2 and PGF2a are BAT-secreted insulin sensitizers and browning inducers;PGE2 and PGF2a sensitize insulin responses through PGE2-EP-pAKT and PGF2a-FP-AKT axis;METTL14-mediated m6A installation selectively destabilizes prostaglandin synthases and their regulator transcripts;Targeting METTL14 in BAT has therapeutic potential to enhance systemic insulin sensitivityAbstract Figure
Evolution of the spider homeobox gene repertoire by tandem and whole genome duplication
Authors: Aase-Remedios Madeleine E. , Janssen Ralf , Leite Daniel J. , Sumner-Rooney Lauren , McGregor Alistair P. 
Published: May 2023

ABSTRACTGene duplication generates new genetic material that can contribute to the evolution of gene regulatory networks and phenotypes. Duplicated genes can undergo subfunctionalisation to partition ancestral functions and/or neofunctionalisation to assume a new function. We previously found there had been a whole genome duplication (WGD) in an ancestor of arachnopulmonates, the lineage including spiders and scorpions but excluding other arachnids like mites, ticks, and harvestmen. This WGD was evidenced by many duplicated homeobox genes, including two Hox clusters, in spiders. However, it was unclear which homeobox paralogues originated by WGD versus smaller-scale events such as tandem duplications. Understanding this is key to determining the contribution of the WGD to arachnopulmonate genome evolution. Here we characterised the distribution of duplicated homeobox genes across eight chromosome-level spider genomes. We found that most duplicated homeobox genes in spiders are consistent with an origin by WGD. We also found two copies of conserved homeobox gene clusters, including the Hox, NK, HRO,Irx, and SINE clusters, in all eight species. Consistently, we observed one copy of each cluster was degenerated in terms of gene content and organisation while the other remained more intact. Focussing on the NK cluster, we found evidence for regulatory subfunctionalisation between the duplicated NK genes in the spiderParasteatoda tepidariorumcompared to their single-copy orthologues in the harvestmanPhalangium opilio. Our study provides new insights into the relative contributions of multiple modes of duplication to the homeobox gene repertoire during the evolution of spiders and the function of NK genes.
Mitochondrial calcium signaling mediated transcriptional regulation of keratin filaments is a critical determinant of melanogenesis
Authors: Tanwar Jyoti , Ahuja Kriti , Sharma Akshay , Sehgal Paras , Ranjan Gyan , Sultan Farina , Priya Anshu , Venkatesan Manigandan , Yenamandra Vamsi K , Singh Archana , Madesh Muniswamy , Sivasubbu Sridhar , Motiani Rajender K 
Published: May 2023

AbstractMitochondria are versatile organelles that regulate several physiological functions. Many mitochondria-controlled processes are driven by mitochondrial Ca2+signaling. However, role of mitochondrial Ca2+signaling in melanosome biology remains unknown. Here, we show that pigmentation requires mitochondrial Ca2+uptake.In vitrogain and loss of function studies demonstrated that Mitochondrial Ca2+Uniporter (MCU) is crucial for melanogenesis while the MCU rheostats, MCUb and MICU1 negatively control melanogenesis. Zebrafish and mouse models showed that MCU plays a vital role in pigmentationin vivo. Mechanistically, MCU controls activation of transcription factor NFAT2 to induce expression of three keratins (keratin 5, 7 and 8), which we report as positive regulators of melanogenesis. Interestingly, keratin 5 in turn modulates mitochondrial Ca2+uptake thereby this signaling module acts as a negative feedback loop that fine-tunes both mitochondrial Ca2+signaling and melanogenesis. Mitoxantrone, an FDA approved drug that inhibits MCU, decreases physiological melanogenesis. Collectively, our data demonstrates a critical role for mitochondrial Ca2+signaling in vertebrate pigmentation and reveal the therapeutic potential of targeting MCU for clinical management of pigmentary disorders. Given the centrality of mitochondrial Ca2+signaling and keratin filaments in cellular physiology, this feedback loop may be functional in a variety of other pathophysiological conditions.HighlightsMCU complex mediated mitochondrial Ca2+uptake is a novel regulator of vertebrate pigmentationKeratin filaments bridge mitochondrial Ca2+signaling to melanosome biogenesis and maturationTranscription factor NFAT2 connects mitochondrial Ca2+dynamics to keratins expressionMCU-NFAT2-Keratin 5 signaling module generates a negative feedback loop to maintain mitochondrial Ca2+homeostasis and to ensure optimal melanogenesisInhibiting MCU with mitoxantrone, an FDA approved drug, leads to reduction in physiological pigmentation
Visualizing dynamic competence pili and DNA capture throughout the long axis ofBacillus subtilis
Authors: Zuke Jason D. , Erickson Rachel , Hummels Katherine R. , Burton Briana M. 
Published: May 2023

AbstractThe first step in the process of bacterial natural transformation is DNA capture. Although long-hypothesized based on genetics and functional experiments, the pilus structure responsible for initial DNA-binding had not yet been visualized forBacillus subtilis. Here, we visualize functional competence pili inBacillus subtilisusing fluorophore-conjugated maleimide labeling in conjunction with epifluorescence microscopy. In strains that produce pilin monomers within ten-fold of wild type levels, the median length of detectable pili is 300nm. These pili are retractile and associate with DNA. Analysis of pilus distribution at the cell surface reveals that they are predominantly located along the long axis of the cell. The distribution is consistent with localization of proteins associated with subsequent transformation steps, DNA-binding and DNA translocation in the cytosol. These data suggest a distributed model forB. subtilistransformation machinery, in which initial steps of DNA capture occur throughout the long axis of the cell and subsequent steps may also occur away from the cell poles.ImportanceThis work provides novel visual evidence for DNA translocation across the cell wall duringBacillus subtilisnatural competence, an essential step in the natural transformation process. Our data demonstrate the existence of natural competence associated, retractile pili that can bind exogenous DNA. Furthermore, we show that pilus biogenesis occurs throughout the cell long axis. These data strongly support DNA translocation occurring all along the lateral cell wall during natural competence, wherein pili are produced, bind to free DNA in the extracellular space, and finally retract to pull the bound DNA through the gap in the cell wall created during pilus biogenesis.
The Tgf-β family member Gdf6Y determines the male sex inNothobranchius furzeriby suppressing oogenesis-inducing genes
Authors: Richter Annekatrin , Mörl Hanna , Thielemann Maria , Kleemann Markus , Geißen Raphael , Schwarz Robert , Albertz Carolin , Koch Philipp , Petzold Andreas , Groth Marco , Hartmann Nils , Herpin Amaury , Englert Christoph 
Published: May 2023

ABSTRACTThe short-lived African killifishNothobranchius furzerilives in seasonal freshwater ponds and has evolved remarkable traits to survive in this limited environment. One of those traits is a genetic XX/XY sex-determination system, which ensures an equal distribution of both sexes. Comparisons of female and male genomic sequences identified the Y-chromosomal copy of the TGF-β family membergdf6as the candidate male sex-determining (SD) gene, which was namedgdf6Yin contrast to the X-chromosomal allelegdf6X. CRISPR/Cas9-mediated inactivation ofgdf6YinN. furzeriled to a complete male-to-female sex reversal in XY animals. The homozygous inactivation ofgdf6Xon the other hand led to a detrimental phenotype post-hatching. This phenotype was compensated bygdf6Y, revealing that the latter became the SD gene while retaining at least some of its originalgdf6function.Gdf6Yis expressed in testicular somatic cells already prior to hatching, where it represses the germ cell-intrinsic feminizing genefoxl2l. We have identified components of the TGF-β signaling pathway, especially the inhibitor of DNA binding genesid1/2/3, and the mRNA decay activatorzfp36l2, as Gdf6Y targets. We conclude that Gdf6Y exerts its function as the male sex-determining gene by suppressing female-specific genes in the developing gonad of maleN. furzeri.
Acute ampakines increase voiding function and coordination in a rat model of SCI
Authors: Rana Sabhya , Alom Firoj , Martinez Robert C , Fuller David D , Mickle Aaron D 
Published: May 2023

AbstractNeurogenic bladder dysfunction causes urological complications and reduces the quality of life in persons with spinal cord injury (SCI). Glutamatergic signaling via AMPA receptors is fundamentally important to the neural circuits controlling bladder voiding. Ampakines are positive allosteric modulators of AMPA receptors that can enhance the function of glutamatergic neural circuits after SCI. We hypothesized that ampakines can acutely stimulate bladder voiding that has been impaired due to thoracic contusion SCI. Adult female Sprague Dawley rats received a unilateral contusion of the T9 spinal cord (n=10). Bladder function (cystometry) and coordination with the external urethral sphincter (EUS) were assessed five days post-SCI under urethane anesthesia. Data were compared to responses in spinal intact rats (n=8). The “low impact” ampakine CX1739 (5, 10, or 15 mg/kg) or vehicle (HPCD) was administered intravenously. The HPCD vehicle had no discernable impact on voiding. In contrast, following CX1739, the pressure threshold for inducing bladder contraction, voided volume, and the interval between bladder contractions were significantly reduced. These responses occurred in a dose-dependent manner. We conclude that modulating AMPA receptor function using ampakines can rapidly improve bladder voiding capability at sub-acute time points following contusion SCI. These results may provide a new and translatable method for therapeutic targeting of bladder dysfunction acutely after SCI.
Rapid decline of prenatal maternal effects with age is independent of postnatal environment in a precocial bird
Authors: Vedder Oscar , Tschirren Barbara , Postma Erik , Moiron Maria 
Published: May 2023

AbstractMaternal effects are an important source of phenotypic variation with potentially large fitness consequences, but how their importance varies with the quality of the environment across an individual’s ontogeny is poorly understood. We bred Japanese quail (Coturnix japonica) of known pedigree and experimentally manipulated the quality of offspring diet, to estimate the importance of prenatal maternal effects in shaping variation in body mass from hatching to adulthood. Maternal genetic effects on body mass at hatching were strong, and largely caused by variation in egg mass, but their importance rapidly declined with age. Whereas there was a large effect of diet on growth, this did not affect the decline of maternal effects variance. The importance of additive genetic and residual variance increased with age, with the latter being considerably larger in the poor diet treatment. Hence, we found no evidence for prenatal maternal effect by postnatal environment interactions, and that prenatal maternal effects are rapidly replaced by direct additive genetic and residual effects when offspring start to develop outside the egg. Thereby these results shed new light on the dynamics of the role of maternal versus offspring genes across ontogeny and environments.
Modifications to the SR-Rich Region of the SARS-CoV-2 Nucleocapsid Regulate Self-Association and Attenuate RNA Interactions
Authors: Reardon Patrick N. , Stuwe Hannah , Shah Sahana , Yu Zhen , Hughes Kaitlyn , Barbar Elisar J. 
Published: May 2023

AbstractThe nucleocapsid protein (N) of SARS-CoV-2 is essential for virus replication, genome packaging, and maturation. N is comprised of two folded domains that are separated by a highly conserved, disordered, Ser/Arg-rich linker, and flanked by disordered tails. Using NMR spectroscopy and analytical ultracentrifugation we identify an alpha-helical region in the linker that undergoes concentration dependent self-association. NMR and gel shift assays show that the linker binds viral RNA but this binding is dampened by both phosphorylation and a naturally occurring mutation, whereas in contrast, RNA binding to the full-length protein is not affected. Interestingly, phase separation with RNA is significantly reduced upon phosphorylation but enhanced with the mutation. We attribute these differences to changes in the linker helix self-association which dissociates upon phosphorylation but forms more stable higher order oligomers in the variant. These data provide a structural mechanism for how the linker region contributes to protein-protein interactions, RNA-protein interactions, liquid-liquid phase separation and N protein regulation.
Mechanically-evoked spike responses of pentascolopidial chordotonal organs ofDrosophilalarvae
Authors: Warren Ben , Göpfert Martin C. 
Published: May 2023

AbstractStretch-sensitive ensembles of neurons in insects, known as Chordotonal organs (COs), function in proprioception, the detection of sound and substrate vibrations. Here we characterize the mechanical sensitivity of the lateral pentascolopidial CO (lch5) ofDrosophilalarvae to establish its postulated role in proprioception. We developed a physiologically realistic method to replicate proprioceptive input to lch5 by pulling the apodeme (tendon) to which the tips of the neurons attach. We found that lch5 sensory neurons respond transiently with a short latency to the velocity-component of stretch displacements and the release of stretch (relaxation). The mechanosensory mutantinactive, has a decreased response to mechanical stimuli and a lower overall spontaneous spike rate. Finally, we simulated the input that lch5 receives during crawling and observed spikes coincident with the start, middle and end of each peristaltic body contraction. We provide the first characterization of proprioceptive feedback inDrosophilalarvae and firmly establish the proprioceptive function of lch5 in larval locomotion.Summary statementWe have characterized the spiking responses of the pentascolopidial chordotonal organ of 20 Drosophila larvae to mechanical stimuli and reveal a clear role in proprioception.
Intratumoral administration of recombinant murine interleukin-12 prevents tumor progression and bone invasion
Authors: Kasahara Shun , Uchihashi Toshihiro , Inubushi Toshihiro , Kurioka Kyoko , Sugauchi Akinari , Miyagawa Kazuaki , Kogo Mikihiko , Tanaka Susumu 
Published: May 2023

ABSTRACTBackgroundOral squamous cell carcinoma (OSCC) progression is accompanied by bone invasion. Therefore, maintaining oral function is necessary to regulate tumor progression. Also, interleukin-12 (IL-12), a well-known anti-tumor cytokine, can suppress osteoclast differentiation in vitro. Accordingly, this study evaluated the therapeutic effects of locally administered IL-12 in an immunocompetent mouse model with mandibular bone invasion mimicking clinical features.MethodsWe investigated anti-bone resorption effects using SCCVII subcutaneous and bone invasion models both in immunocompetent and athymic mice. Furthermore, we measured bone resorption using micro-computed tomography.ResultsIntratumoral injection of recombinant murine IL-12 (r-mIL-12) significantly prolonged immunocompetent mouse survival and suppressed tumor growth and bone resorption. Real-time PCR analysis revealed that interferon-gamma (IFN-γ) and Fas ligand (FasL) were upregulated after r-mIL-12 administration, compared to control levels. However, when the athymic mouse bone invasion model was evaluated, r-mIL-12-mediated suppression of tumor growth and bone resorption were equivalent to those observed in the control group, highlighting the key role of T cells in the bone invasion.Conclusionsr-mIL-12 may represent a potent therapeutic agent for OSCC accompanied by bone invasion.SUMMARYIntratumoral injection of recombinant murine IL-12 showed anti-tumor and anti-bone resorption effects in an immunocompetent mouse bone invasion model through a T cell-dependent mechanism.
BIG participates in the Arg/N-degron pathways and the hypoxia response inArabidopsis thaliana
Authors: Zhang Hongtao , Rundle Chelsea , Winter Nikola , Miricescu Alexandra , Mooney Brian C. , Bachmair Andreas , Graciet Emmanuelle , Theodoulou Frederica L. 
Published: May 2023

AbstractBIG (also known as DOC1 and TIR3) is an 0.5 MDa protein that has been associated with multiple important functions in signalling and development through forward genetic screens inArabidopsis thaliana. However, the biochemical function(s) of BIG are unknown. Here, we investigated whether BIG plays a role in the Arg/N-degron pathways, protein regulatory mechanisms in which substrate protein fate is influenced by the N-terminal (Nt) residue. In Arabidopsis, PROTEOLYSIS1 (PRT1) is an E3 ligase with specificity for aromatic amino acids, whereas PROTEOLYSIS6 (PRT6) targets basic N-terminal residues. We crossed a big loss-of-function allele toprt6andprt1mutants and examined the stability of protein substrates. Stability of model N-degron pathway substrates was enhanced inprt6-1 big-2andprt1-1 big-2relative to the respective single mutants. Abundance of the PRT6 physiological substrates, HYPOXIA RESPONSIVE ERF (HRE)2 and VERNALIZATION (VRN)2 was similarly increased inprt6 bigdouble mutants, without increase in transcripts. Accordingly, hypoxia marker expression was enhanced inprt6 bigdouble mutants, in a manner requiring arginyltransferase activity and RAP-type ERFVII transcription factors. Transcriptomic analysis of roots not only demonstrated synergistically increased expression of a plethora of hypoxia responsive genes in the double mutant relative toprt6but also revealed other roles for PRT6 and BIG, including regulation of suberin deposition through both ERFVII-dependent and independent mechanisms, respectively. Our results show that BIG acts together with PRT6 to regulate the hypoxia response and wider processes.Significance StatementThe N-degron pathways are a group of protein regulatory mechanisms that play important roles in plant growth, development, and response to biotic and abiotic stresses. Despite rapid progress in the last decade, key enzymatic components of the pathways remain to be identified. BIG (also known as DOC1 and TIR3) is a protein of approximately 0.5 MDa, associated with multiple, distinct roles in plants but the precise biochemical functions of this protein have remained enigmatic until now. Here we identify BIG as a new component of plant N-degron pathways that acts together with the N-recognin E3 ligase PROTEOLYSIS6 (PRT6) to control the hypoxia response and other functions inArabidopsis thaliana.
GPI-anchored Gas1 protein regulates cytosolic proteostasis in yeast
Authors: Wang Yuhao , Ruan Linhao , Li Rong 
Published: May 2023

AbstractDecline in protein homeostasis (proteostasis) is a hallmark of cellular aging and aging-related diseases. Maintaining a balanced proteostasis requires a complex network of molecular machineries that govern protein synthesis, folding, localization, and degradation. Under proteotoxic stress, misfolded proteins that accumulate in cytosol can be imported into mitochondria for degradation via ‘mitochondrial as guardian in cytosol’ (MAGIC) pathway. Here we report an unexpected role of yeast Gas1, a cell wall-bound glycosylphosphatidylinositol (GPI)-anchored β-1,3-glucanosyltransferase, in differentially regulating MAGIC and ubiquitin-proteasome system (UPS). Deletion of Gas1 inhibits MAGIC but elevates polyubiquitination and UPS-mediated protein degradation. Interestingly, we found that Gas1 exhibits mitochondrial localization attributed to its C-terminal GPI anchor signal. But this mitochondria-associated GPI anchor signal is not required for mitochondrial import and degradation of misfolded proteins via MAGIC. By contrast, catalytic inactivation of Gas1 via thegas1E161Qmutation inhibits MAGIC but not its mitochondrial localization. These data suggest that the glucanosyltransferase activity of Gas1 is important for regulating cytosolic proteostasis.
Extended Nucleic Acid (exNA): A Novel, Biologically Compatible Backbone that Significantly Enhances Oligonucleotide Efficacyin vivo
Authors: Yamada Ken , Hariharan Vignesh Narayan , Caiazzi Jillian , Miller Rachael , Furguson Chantal , Sapp Ellen , Fakih Hassan , Tan Qi , Yamada Nozomi , Furgal Raymond C. , Paquette Joseph , Bramato Brianna , McHugh Nicholas , Summers Ashley , Lochmann Clemens , Godinho Bruno M.D.C. , Hildebrand Samuel , Echeverria Dimas , Hassler Matthew R. , Alterman Julia F. , DiFiglia Marian , Aronin Neil , Khvorova Anastasia 
Published: May 2023

AbstractMetabolic stabilization of therapeutic oligonucleotides requires both sugar and backbone modifications, where phosphorothioate (PS) is the only backbone chemistry used in the clinic. Here, we describe the discovery, synthesis, and characterization of a novel biologically compatible backbone, extended nucleic acid (exNA). Upon exNA precursor scale up, exNA incorporation is fully compatible with common nucleic acid synthetic protocols. The novel backbone is orthogonal to PS and shows profound stabilization against 3’- and 5’-exonucleases. Using small interfering RNAs (siRNAs) as an example, we show exNA is tolerated at most nucleotide positions and profoundly improves in vivo efficacy. A combined exNA-PS backbone enhances siRNA resistance to serum 3’-exonuclease by ∼32-fold over PS backbone and >1000-fold over the natural phosphodiester backbone, thereby enhancing tissue exposure (∼6-fold), tissues accumulation (4- to 20-fold), and potency both systemically and in brain. The improved potency and durability imparted by exNA opens more tissues and indications to oligonucleotide-driven therapeutic interventions.