Neutrophil extracellular traps (NETs) are increasingly recognized as key regulators of tumor progression, yet the molecular circuitry that governs their induction in cancer remains elusive. Here, we identify the RNA-bind...Neutrophil extracellular traps (NETs) are increasingly recognized as key regulators of tumor progression, yet the molecular circuitry that governs their induction in cancer remains elusive. Here, we identify the RNA-binding protein RBFOX2 as a tumor suppressor that curtails glioma growth by coordinately restraining tumor cell proliferation and NETosis. RBFOX2 expression is markedly reduced in glioma and positively correlates with patient survival. Mechanistically, RBFOX2 binds to 5-hydroxymethylcytidine (5hmC)-modified sites within PDGFB mRNA and promotes its decay, thereby dampening AKT-SP1 signaling and repressing CSF3 transcription. This repression limits neutrophil-mediated NET formation in the tumor microenvironment, as confirmed in PAD4 mice and upon CSF3 neutralization. Collectively, our study uncovers a 5hmC-dependent post-transcriptional mechanism linking RBFOX2 to NETosis control and glioma suppression, revealing RBFOX2 as a potential biomarker and therapeutic lever and establishing a broader paradigm in which RNA-binding proteins couple post-transcriptional RNA modification and immune regulation in tumor evolution.
Mutant KRAS-driven control of protein synthesis remains poorly defined. Here, we define KRAS-dependent translational programs and their acute remodeling upon KRAS inhibition. We find that mutant KRAS controls the transla...Mutant KRAS-driven control of protein synthesis remains poorly defined. Here, we define KRAS-dependent translational programs and their acute remodeling upon KRAS inhibition. We find that mutant KRAS controls the translation of a subset of mRNAs and affects the production of proteins of the mRNA translation apparatus. Interestingly, these specific subsets of mRNAs have short, weakly folded 5'UTRs and harbor low folding energy consensus RNA sequences. We observe ribosome accumulation on selective mRNAs. Our findings clarify the indispensable role of mutant KRAS in regulating mRNA translation, setting it apart from the other previously known mechanisms that depend on mTOR and EIF4E-EIF4A signals. Our findings uncover a mechanism by which mutant KRAS selectively uncouples the translation of mRNAs for protein synthetic machinery from the broader mRNA pool, redefining our understanding of the oncogenic regulation of mRNA translation in cancer.
The nuclear pore complex (NPC) basket has been implicated in regulating meiotic recombination, but the underlying mechanism remained elusive. Here, we show that most basket subunits are required for controlled crossing-o...The nuclear pore complex (NPC) basket has been implicated in regulating meiotic recombination, but the underlying mechanism remained elusive. Here, we show that most basket subunits are required for controlled crossing-over in budding yeast. Central to this function, the nucleoporin Nup60 anchors the SUMO protease Ulp1 at the nuclear periphery, thereby protecting the synaptonemal complex (SC) protein Ecm11 from premature deSUMOylation. Unscheduled dissociation of Ulp1 from the NPC impairs Ecm11 SUMOylation, disrupts synapsis, elevates crossovers, and compromises gamete viability. Remarkably, engineered tethering of Ulp1 to the NPC restores SC integrity and recombination control in basket mutants. We further show that Polo-like kinase Cdc5 remodels SUMO homeostasis at the prophase I-metaphase I transition, triggering partial Ulp1 release from the NPC and phosphorylating the SUMO ligases Siz1 and Siz2. These findings uncover how nucleoporins, SUMO enzymes, and kinase signaling cooperate to coordinate SC dynamics with crossover control, safeguarding meiotic genome transmission.
Plucińska K, Wayne CR, Sanford H
… +11 more, Mathew B, Ropek N, Adaniya SM, Model C, Gómez-Banoy N, Morozova K, Cao X, Friedman JM, Loh KH, Cohen P, Vinogradova EV
Intercellular communication is critical for maintaining organismal metabolic homeostasis. Here, we develop a method enabling temporally controlled, cell-type-specific labeling of secreted and membrane proteins in key met...Intercellular communication is critical for maintaining organismal metabolic homeostasis. Here, we develop a method enabling temporally controlled, cell-type-specific labeling of secreted and membrane proteins in key metabolic tissues. The method employs a genetically encoded proximity-labeling strategy by targeting a Cre-dependent TurboID ligase to the endoplasmic reticulum (ER) in ES cell-derived mice. The expression of TurboID in hepatocytes, adipocytes, and B lymphocytes enabled the characterization of cell type-specific ER proteomes at baseline and in response to fasting, inflammation, and dietary obesity, revealing tissue- and perturbation-specific changes and augmenting our understanding of how the proteomes of individual tissues change to regulate systemic energy balance. This comprehensive resource represents an important advance toward understanding both how cell-to-cell communication changes in response to energy balance and how it contributes to these alterations. This method is broadly applicable and provides a means for identifying biomarkers and therapeutic targets across a wide range of tissues.
Systemic lupus erythematosus (SLE) is an autoimmune disease preferentially observed in women. X-linked gene expression in XX females is equalized with that in XY males by X chromosome inactivation (XCI). However, B cells...Systemic lupus erythematosus (SLE) is an autoimmune disease preferentially observed in women. X-linked gene expression in XX females is equalized with that in XY males by X chromosome inactivation (XCI). However, B cells from female patients with SLE and mouse models of SLE exhibit aberrant X-linked gene expression and mislocalization of Xist RNA, a critical regulator of XCI, suggesting that impaired XCI may contribute to the disease. Here, some female mice harboring a B cell-specific Xist deletion ("Xist cKO") spontaneously developed SLE phenotypes, including expanded activated B cell subsets, disease-specific autoantibodies, and glomerulonephritis. Pristane treatment of Xist cKO mice led to an increased expansion of activated B cell subsets, resulting in higher anti-dsDNA autoantibody production. Activated B cells from Xist cKO mice with SLE phenotypes had an increased expression of X-linked and autosomal genes, depletion of X-linked H3K27me3, and were more responsive to stimulation. This work suggests that impaired XCI maintenance in B cells directly contributes to SLE-associated phenotypes in a female-biased manner.
Sepsis is a life-threatening condition characterized by infection-induced organ dysfunction, with fluid imbalance and cardiovascular instability as cardinal features. Although circulating procalcitonin (PCT) is widely us...Sepsis is a life-threatening condition characterized by infection-induced organ dysfunction, with fluid imbalance and cardiovascular instability as cardinal features. Although circulating procalcitonin (PCT) is widely used as a diagnostic and prognostic marker in sepsis, its pathophysiological role remains poorly understood. Here, we identify a central neural circuit through which PCT directly disrupts fluid homeostasis: systemic PCT crosses the blood-brain barrier, activates calcitonin receptors, and depolarizes the Oprk1-expressing neurons in the ventromedial preoptic nucleus of the hypothalamus (VMPO). In vivo, PCT administration induces polydipsia and polyuria-phenotypes recapitulated by chemogenetic stimulation of VMPO neurons. We demonstrate that VMPO neurons project to and activate arginine vasopressin (AVP)-expressing neurons in the supraoptic nucleus (SON), leading to increased blood pressure. Together, our findings define a PCT-sensitive VMPO→SON neural circuit that integrates fluid balance and cardiovascular regulation. Our data highlight critical role of the brain in coordinating organ pathophysiology during infection.
The periosteum, a thin membranous tissue covering the external bone surface, is a well-established source of osteogenic activity during bone growth and fracture repair. However, the molecular identity and anatomical nich...The periosteum, a thin membranous tissue covering the external bone surface, is a well-established source of osteogenic activity during bone growth and fracture repair. However, the molecular identity and anatomical niche of its progenitor cells remain incompletely defined. Using single-cell RNA sequencing and RNA in situ hybridization in mouse long bones, we identified that fibroblast growth factor 18 (Fgf18) expression marks a discrete subset of cells within the fibrous layer of the periosteum that co-express genes associated with skeletal stem and progenitor populations. Lineage tracing revealed that Fgf18 cells contribute minimally to cortical bone formation under homeostatic conditions but robustly expand and generate chondrocytes and osteoblasts following injury. Targeted ablation of Fgf18-lineage cells impaired fracture healing, confirming their functional importance. Together, these findings define Fgf18 fibrous periosteal cells as a reserve progenitor population that is activated by injury to regenerate bone.
Orthomarburgviruses are the etiological agents of human Marburg virus disease, with case fatality rates reaching up to 90%. Of the limited known targets for protective antibodies on the orthomarburgvirus surface glycopro...Orthomarburgviruses are the etiological agents of human Marburg virus disease, with case fatality rates reaching up to 90%. Of the limited known targets for protective antibodies on the orthomarburgvirus surface glycoprotein (GP), the GP2 wing region is undefined structurally, and its recognition by antibodies is poorly understood. We report a comprehensive genetic and structural analysis of four independent broadly reactive antibody lineages that target a common epitope within the GP2 wing, including macaque antibodies CM10, CM11.1, and CM12.1 and murine antibody 30G4. Co-crystal structures reveal the recognition of conserved structural determinants within the wing by all four antibodies. Immunogenetic analyses uncover common antibody sequence signatures that underlie interactions with the wing, leading in some cases to nearly identical modes of antibody binding. Our study advances the structural understanding of the GP2 wing protective region and defines conserved immunogenetic and structural features for broad antibody recognition, informing the development of vaccines and antibody countermeasures.
Natural killer (NK) cells are cytotoxic innate lymphoid cells which directly kill tumor cells, thus represent an attractive target for immunotherapy. However, NK cells face immunosuppression in the tumor microenvironment...Natural killer (NK) cells are cytotoxic innate lymphoid cells which directly kill tumor cells, thus represent an attractive target for immunotherapy. However, NK cells face immunosuppression in the tumor microenvironment (TME), rendering them dysfunctional. While cancer-associated fibroblasts (CAFs) represent an abundant, heterogeneous component of pancreatic ductal adenocarcinoma (PDAC), their interplay with NK cells is largely understudied. Analyzing human samples and employing mouse models of PDAC and functional assays, we observed that intratumoral NK cells are immature, and TGF-β driven myofibroblastic (my)CAFs are strong NK suppressors, in contrast to inflammatory (i)CAF. Furthermore, myCAF-enriched tumor areas excluded NK cells, consistent with their reduced capacity to attract NK cells. Pancreatic CAFs in general reduced NK cell cytotoxicity by direct contact and via soluble factors, including prostaglandin E2 (PGE2). This work reveals distinct and overlapping roles of CAF subpopulations on NK cell functions, suggesting that overcoming CAF-imposed barriers to NK cytotoxicity and tumor infiltration is essential to unleash their anti-tumoral properties.
Olinger B, Anerillas C, Herman AB
… +12 more, Tsitsipatis D, Banarjee R, Tanaka T, Candia J, Maragkakis M, Bandinelli S, Walker KA, Simonsick EM, Qi YA, Ferrucci L, Gorospe M, Basisty N
Cellular senescence is implicated in age-related pathologies, and identifying circulating biomarkers of senescence holds great diagnostic potential. Circulating senescence signatures are predictive of age-related traits...Cellular senescence is implicated in age-related pathologies, and identifying circulating biomarkers of senescence holds great diagnostic potential. Circulating senescence signatures are predictive of age-related traits and diseases, though cell type senescence signatures have not been comprehensively explored. In this study, senescence signatures from the Senescence Catalog (SenCat), including 14 human cell types are examined in circulation for clinical relevance in two longitudinal studies-1,275 participants of the Baltimore Longitudinal Study of Aging (BLSA) and 997 participants of the Invecchiare in Chianti (InCHIANTI) study. Notably, pooled senescence proteins outperform non-senescence proteins in predicting many clinical parameters such as age and hypertension, and in many instances, cell type senescence signatures map most strongly to their corresponding health domain. Importantly, the immune cell senescence signature is associated with mortality and future disease onset. This study demonstrates that circulating cell type biomarkers of senescence can reveal higher resolution health status than previously attained.
The decoding center (DC) is a key ribosomal structure for accurate translation, assembled in a multi-step process that starts on nucleolar pre-ribosomes and ends in the cytoplasm. While late cytoplasmic steps and their c...The decoding center (DC) is a key ribosomal structure for accurate translation, assembled in a multi-step process that starts on nucleolar pre-ribosomes and ends in the cytoplasm. While late cytoplasmic steps and their checkpoint mechanisms are well characterized, the regulation of early nucleoplasmic DC assembly is unclear. Here, we show that the essential assembly factor Rrp12 plays a central coordinating role. Using Chaetomium thermophilum and cryo-electron microscopy analyses of fifteen pre-40S intermediates, we demonstrate that Rrp12 C terminus truncation: (1) inhibits release of the Utp14-Dhr1 pair, (2) displaces Tsr1, (3) promotes premature stabilization of h28, and (4) prevents h44 formation. These defects impair final 18S rRNA processing and prematurely activate the quality control kinase Rio1. Our results reveal a nucleoplasmic checkpoint during DC formation and establish Rrp12 as a critical regulator ensuring accurate assembly and orderly ribosome maturation.
McCune A, Usha MK, Baldus I
… +10 more, Willett CJ, Ferrito N, Barakat R, Clemensen E, Snella E, Morton M, Lacal J, Pavani G, Campbell CA, Espin-Palazon R
Perturbations in cell fate disrupt tissue homeostasis and drive diseases such as cancer. The JAK/STAT pathway is central to hematopoietic malignancies, yet its regulators in cell fate decisions remain poorly defined. Her...Perturbations in cell fate disrupt tissue homeostasis and drive diseases such as cancer. The JAK/STAT pathway is central to hematopoietic malignancies, yet its regulators in cell fate decisions remain poorly defined. Here, we identify progranulin (GRN) as a critical determinant of myeloid fate through the regulation of JAK2/STAT3 signaling. Using inducible stat3 and grna zebrafish models, knockout approaches, FACS, transcriptomics, and CUT&RUN, we show that Stat3 induces grna, which in turn amplifies stat3 expression, reinforcing myeloid over erythroid commitment. Lineage tracing and live imaging reveal that this Grna/Stat3 feedback operates during definitive, but not primitive, myelopoiesis. Functionally, definitive myeloid cells drive tissue repair, whereas primitive counterparts are less effective. Myeloid differentiation in a human leukemia line required GRN co-stimulation of JAK2/STAT3, highlighting its conserved role in fate determination. These findings uncover a regulatory axis controlling hematopoietic commitment and identify GRN as a potential therapeutic target in diseases with aberrant JAK2/STAT3 activation.
Touch is essential for survival, guiding interactions with the environment and forming the basis of social and emotional connection. While touch perception is mediated by mechanoreceptors composed of neurons and glia, th...Touch is essential for survival, guiding interactions with the environment and forming the basis of social and emotional connection. While touch perception is mediated by mechanoreceptors composed of neurons and glia, the role of glia in mechanosensation remains poorly understood. Here, we uncover a previously unrecognized function for glia-derived serotonin in regulating touch sensitivity in Caenorhabditis elegans. Using in vivo Ca and serotonin imaging, fluorescence microscopy, and behavioral assays, we show that amphid sheath (AMsh) glia release serotonin via dense core vesicles. This serotonin activates the neuronal receptor SER-5 to enable ASH neuron responses, while simultaneously engaging glial MOD-1 autoreceptors to provide feedback control. We further identify a concentration-dependent mechanism that fine-tunes sensory sensitivity through cAMP- and Ca-dependent pathways. Together with prior evidence of glial GABA release, our findings establish glia as active regulators of touch, balancing excitation and inhibition to maintain mechanosensory function.
Increasing cell size drives proteomic changes that impact cell physiology. However, the molecular basis of size-dependent proteome remodeling has remained unclear. Here, we develop an inducible Cyclin D1 expression syste...Increasing cell size drives proteomic changes that impact cell physiology. However, the molecular basis of size-dependent proteome remodeling has remained unclear. Here, we develop an inducible Cyclin D1 expression system in human cells to generate proliferating cells spanning over a 2-fold size range. We use this system to make comprehensive genome-wide measurements of mRNA and protein concentrations and stability. We find that protein and mRNA turnover rates are weakly related to cell size but that mRNA concentrations are strongly size-dependent. This establishes that transcriptional regulation is the basis of proteome remodeling. Live-cell imaging of nascent mRNAs using the MS2 system is used to measure how transcriptional dynamics change with cell size. Larger cells prolong transcriptional bursts but maintain similar burst amplitudes to achieve transcriptional scaling. Together, our results show how transcription is modulated by cell size to remodel the proteome and alter cell physiology.
Eukaryotic DNA wraps around histone octamers forming nucleosomes, which modulate genome function by defining chromatin environments with distinct accessibility. These well-conserved properties allowed "humanization" of t...Eukaryotic DNA wraps around histone octamers forming nucleosomes, which modulate genome function by defining chromatin environments with distinct accessibility. These well-conserved properties allowed "humanization" of the nucleosome core particle (NCP) in Saccharomyces cerevisiae at high fitness costs. Here, we studied histone-humanized yeast genomes to understand how species-specific chromatin affects nuclear organization and function. We found a size increase in human-NCP, linked to shorter free linker DNA, supporting decreased chromatin accessibility. Three-dimensional (3D) humanized genome maps showed increased chromatin compaction and defective centromere clustering, correlated with high chromosomal aneuploidy rate. Site-specific chromatin alterations were associated with lack of initiation of early origins of replication and dysregulation of the ribosomal (rDNA and rRNA) metabolism. This latter led to nucleolar fragmentation and rDNA-array instability, through a non-coding RNA-dependent mechanism, leading to its extraordinary, but entirely reversible, intra-chromosomal expansion. Overall, our results reveal species-specific properties of the NCP that define epigenome function across vast evolutionary distances.
The activity of many oncogenic transcription factors (TFs) is constrained by enhancer binding-site affinity, leaving many low-affinity sites unoccupied under physiological conditions. Whether coactivator amplification in...The activity of many oncogenic transcription factors (TFs) is constrained by enhancer binding-site affinity, leaving many low-affinity sites unoccupied under physiological conditions. Whether coactivator amplification in cancer redirects TFs to these sites is unclear. Here, we show that amplification of MED30, a Mediator subunit at 8q24, promotes aberrant MYC binding to low-affinity regulatory regions and is associated with poor outcomes. Besides frequent MYC MED30 co-amplification, MED30 overexpression alone is sufficient to enable MYC occupancy and activation of previously weak or unbound enhancers and promoters, driving tumor-promoting gene expression. Functional studies in pancreatic ductal adenocarcinoma and glioblastoma demonstrate that MED30 is oncogenic and serves as a prognostic marker independent of MYC amplification. These findings reveal a cofactor-driven mechanism by which MED30 licenses MYC binding to low-affinity sites, reprogramming enhancers during cancer progression with therapeutic implications.
Similarities and differences in the self-assembly of actin filaments from different species inform our understanding of its evolution. However, this basic knowledge is largely incomplete. Here, we systematically characte...Similarities and differences in the self-assembly of actin filaments from different species inform our understanding of its evolution. However, this basic knowledge is largely incomplete. Here, we systematically characterize assembly kinetics for actin from two yeast species that are five hundred million years apart in evolution, Saccharomyces cerevisiae and Schizosaccharomyces pombe, and compare them to the well-studied rabbit muscle actin from which they diverged a billion years ago. We find that, in the ATP state, both yeast actins behave strikingly like mammalian actin at filament barbed ends. In contrast, yeast actin filaments in both the ADP·Pi and the ADP states depolymerize several-fold faster than their mammalian counterparts, and they release inorganic phosphate over 20-fold faster. We show that the absence of methylation on histidine 73 largely accounts for this faster aging of yeast actin filaments. We also reveal biochemical and mechanical differences between the actins of the two yeasts. Our findings suggest that actins are more diverse and biochemically specialized across species than previously recognized.
Temperature has a severe impact on maize height. However, the mechanisms by which maize regulates height in response to heat have not been fully established. Here, we perform a genome-wide association study and identify...Temperature has a severe impact on maize height. However, the mechanisms by which maize regulates height in response to heat have not been fully established. Here, we perform a genome-wide association study and identify one single nucleotide polymorphism (SNP) located in the maize MIR172C promoter strongly associated with temperature-dependent plant height. This SNP is selected during maize domestication from tropical to temperate zones. Molecular evidence shows that the G-to-A variation enhances the binding of PIF4 to the MIR172C, greatly represses MIR172C transcription. Phenotyping indicates that MIR172C is highly expressed in the meristem to promote internode elongation. miR172c represses its target IDS1 and activates the brassinosteroid (BR) pathway in a temperature-dependent manner. Under heat stress, miR172s regulate IDS1 predominantly by translational inhibition. This study identifies a natural variation modulating temperature-dependent maize height and elucidates the underlying mechanism. This finding reports a key locus for future breeding to enhance environmental adaptability in maize.
Chemoresistance is a primary factor limiting nasopharyngeal carcinoma (NPC) treatment. Growing evidence indicates that E3 ubiquitin ligases play a pivotal role in chemoresistance. Here, we identified that the E3 ubiquiti...Chemoresistance is a primary factor limiting nasopharyngeal carcinoma (NPC) treatment. Growing evidence indicates that E3 ubiquitin ligases play a pivotal role in chemoresistance. Here, we identified that the E3 ubiquitin ligase RNF138 is significantly upregulated in NPC patients who do not respond to chemotherapy. Our study reveals that RNF138 promotes the K48-linked ubiquitination of hnRNPA0 at K133, thereby destabilizing WWOX mRNA. The subsequent loss of WWOX protein relieves the inhibition of JAK2 self-phosphorylation, leading to constitutive pathway activation. Consequently, RNF138-JAK2/STAT3 activation suppresses chemotherapy-induced apoptosis via reduced ROS production and promotes immune evasion by upregulating PD-L1. Clinically, high RNF138 expression correlated with poor prognosis and resistance to chemotherapy. In conclusion, this study unveils the RNF138-hnRNPA0-WWOX axis as a driver of JAK2/STAT3 activation, leading to both chemoresistance and immune evasion in NPC. This work positions RNF138 as a valuable biomarker to guide individualized chemotherapy, and highlights JAK inhibitors as a potential targeted therapy for NPC patients.
During meiosis I, the cohesin Rec8 is cleaved by separase along the chromosome arms but is protected at the centromere by shugoshin (Sgo1); during meiosis II, it is not protected. In fission yeast, another meiotic regula...During meiosis I, the cohesin Rec8 is cleaved by separase along the chromosome arms but is protected at the centromere by shugoshin (Sgo1); during meiosis II, it is not protected. In fission yeast, another meiotic regulator, meikin (Moa1), supports the protective function of Sgo1 primarily by phosphorylating Rec8 at S450. Here we show that both meiosis I-specific proteins, Sgo1 and Moa1, are degraded during anaphase I by the APC/C-Slp1 pathway. To explore the possibility of ectopic protection during meiosis II, we expressed non-degradable forms of Moa1 and Sgo1 during meiosis. Our analyses revealed that stabilization of the Sgo1 protein and phosphorylation of Rec8 at S449 and S450 are necessary and sufficient for the protection of Rec8 cohesin during meiosis II. Furthermore, our results suggest that the phosphorylation-dependent interaction between Rec8 and Sgo1 during meiosis is prominent at mono-oriented kinetochores but not at bi-oriented kinetochores.