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Hernandez, C.

Publications and source records attributed to Hernandez, C..

At least 19 recordsLinked to original sources

The Signaling Pathway of a Bacterial Two-Component System Determined by cryo-EM

Although two-component systems (TCSs) play ubiquitous roles in sensing signals for bacterial survival and virulence, a detailed structural map of the signal transduction pathway is lacking. Bacteriophytochromes (BphPs) are enzymatic photoreceptors with homologs in plants that regulate bacterial physiological responses via TCS signaling. Here, we employ cryo-EM to elucidate the structural basis of a light-regulated TCS by capturing the full-length BphP histidine kinase (HK) in complex with its downstream response regulator (RR). The RR binds as a dimer to the HK but contacts the enzyme only through one subunit. Light absorption perturbs the entire BphP structure, including the HK domain, which disrupts phosphorylation and phosphotransfer reactions and promote TCS complex dissociation. Thus, light controls signaling through both HK activity and reversible binding of the RR.

biophysics↗

Life history traits predict the contribution of transient dynamics to variation in population growth

The performance of any natural population in variable environments depends on contemporaneous changes in its vital rates (e.g., survival, reproduction) as well as legacies carried by its population structure. Yet whether the relative contribution of these two pathways can be predicted from life history remains unknown. Here, we use stochastic simulations of 1,986 matrix population models from 137 species to quantify the contribution of transient dynamics to variation in population growth rate, and test its associations with key life history traits. Longer generation times were associated with reductions in transient contributions, contrary to theoretical expectations. Greater stage-specific survival heterogeneities were associated with increases in transient contributions, whereas greater iteroparity was associated with decreases in plants but increases in animals. These associations were robust to body size, phylogenetic relationships, and vital-rate variability. Life history traits therefore provide a strong predictor for when population structure shapes population responses to environmental variability.

ecology↗

Microbial exometabolite responses to root exudates from tallgrass prairie plants

Root exudates influence soil microbial community assembly and function, yet plant species-specific effects on microbially-driven metabolic transformations remain insufficiently characterized under controlled conditions. Here, we present a multi-omic dataset of exudate-microbe interactions across six plant species, including tallgrass prairie species (Helianthus pauciflorus, Carex brevior, Eragrostis spectabilis, Astragalus canadensis, and Panicum virgatum) and a model grass (Brachypodium distachyon), generated using an in vitro incubation experiment. Sterile root exudates were used to amend a soil-mimicking medium inoculated with a native tall-grass prairie soil microbiome. The dataset includes full-length 16S rRNA gene sequencing for microbial community profiling, untargeted LC-MS/MS metabolomics for exometabolomic profiling, and optical density measurements of microbial growth. We describe the axenic plant growth protocols, experimental design, data acquisition, processing workflows, and technical validation. This dataset provides a resource for investigating microbially mediated exometabolite transformations across diverse plant species to help understand rhizosphere processes, microbiome assembly, and ecosystem function.

microbiology↗

Epistasis of two classical color genes, B and L-2, synergistically controls carotenoid accumulation in squash

Carotenoid accumulation underlies fruit color and nutritional quality in squash (Cucurbita pepo). One pair of dominant genes, B and L-2, have been long known to interact epistatically, substantially boosting carotenoid accumulation and producing intensely orange-fleshed fruit. However, their molecular identities and regulatory mechanism are unknown. Here, we show that B encodes a truncated H subunit of magnesium chelatase (CpCHLHB) and L-2 encodes a homolog of Arabidopsis Pseudo-Response Regulator 2 (CpAPRR2-A). Significantly, expression of phytoene synthase (CpPSY-A), which encodes the major rate-limiting enzyme in carotenoid biosynthesis, was dramatically upregulated in fruit of B/B L-2/L-2 plants compared with b/b L-2/L-2 or B/B l-2/l-2, showing that the B and L-2 interaction affects CpPSY-A transcription. A similar upregulation was also observed in Arabidopsis gun5 L-2 transgenic plants, where gun5 is a genetic mimic of the C. pepo B gene. The wild-type CpCHLHb physically interacted with CpAPRR2-A, attenuating the CpAPRR2-A-mediated activation of CpPSY-A. In contrast, the truncated CpCHLHB lost its ability to interact with CpAPRR2-A, enabling CpAPRR2-A to activate CpPSY-A and produce intensely orange fruit. These findings uncover the mechanism underlying the epistatic interaction through which B and L-2 act synergistically to boost carotenoid production, offering novel mechanistic insights and key targets for improving crop quality. One-sentence summarySynergistic epistasis between B and L-2 arises from loss of interaction between their encoded proteins, resulting in dramatically upregulating the key rate-limiting enzyme in carotenoid biosynthesis pathway to produce intensely orange-fleshed fruit in squash.

plant biology↗

Stage-dependent biotic interactions may not be important for stochastic competitive dynamics with little variation in stage structure

O_LIStage-dependent interactions, in which different life cycle stages (e.g., juveniles, adults) exert different per-capita competitive effects, are widespread across ecological communities. However, whether explicitly accounting for such ontogenetic variation improves forecasts of stochastic community dynamics remains unclear. We tested how the strength of stage dependence and species life-history strategy influence the predictive accuracy of community models that either include or ignore stage-specific interactions. C_LIO_LIWe constructed stochastic two-species competition models using stage-structured matrix population models spanning five virtual life histories along the fast-slow continuum. Density dependence was imposed separately on juvenile survival, adult survival, progression, retrogression, or fertility, and the strength of stage dependence varied from adult-driven to juvenile-driven competition. We then fitted deterministic projection models with and without stage-dependent interaction terms to simulated time series and quantified predictive performance over 100 time-step forecasts using mean absolute percentage error (MAPE). C_LIO_LIIncreasing stage dependence consistently reduced the predictive accuracy of models that ignored stage structure. However, absolute prediction errors remained small across all scenarios (MAPE < 0.7%), even under strong stage dependence. The influence of life-history strategy depended on which vital rate was density dependent: when juvenile survival was density dependent, faster life histories showed larger errors; when progression, retrogression, or fertility were density dependent, slower life histories exhibited greater errors; and when adult survival was density dependent, no consistent life-history effect emerged. Across simulations, temporal variation in population structure was low (coefficient of variation < 0.036), and prediction error was strongly associated with the magnitude of structural fluctuations rather than life-history pace per se. C_LIO_LISynthesis. Stage-dependent interactions can, in principle, alter stochastic competitive dynamics, but their practical importance for ecological forecasting depends on the extent to which population stage structure fluctuates through time. When environmental stochasticity dominates and stage structure remains near equilibrium, simpler models that ignore stage dependence provide robust approximations of community dynamics. Our results identify conditions under which demographic detail is necessary for forecasting and highlight the central role of structural variability in linking life-history strategy to community-level dynamics. C_LI

ecology↗

Standing genetic variation underlies divergence of nicotinic acetylcholine receptor subunits among cryptic species of the Anopheles gambiae complex

Arthropod species differ in insecticide susceptibility, yet how pre-existing polymorphism at target sites shapes variable responses within and among populations remains poorly understood. Recently diverged taxa provide ideal systems to test how target-site divergence modulates species sensitivity. Using whole-genome sequencing data from 573 mosquitoes representing six cryptic species of the Anopheles gambiae complex, we analyzed standing genetic variation across all 11 nicotinic acetylcholine receptor (nAChR) subunit genes to establish a baseline for natural diversity before the large-scale deployment of nAChR-targeting insecticides in Africa. We detected no previously reported resistance alleles from agricultural pests and found no evidence of selective sweeps or loss-of-function mutations across the nAChR gene family. Patterns of polymorphism were consistent with strong purifying selection. Most nonsynonymous variants were rare, predicted to be tolerated by SIFT (score [&ge;] 0.05), present almost exclusively in heterozygotes, and occurred outside ligand-binding and transmembrane domains. However, the 6 subunit exhibited relaxed constraint, with two high-frequency substitutions (I198M and D202E) that defined haplotypes segregating by species. The derived alleles represented ancient polymorphisms, showed evidence of introgression, and were fixed in populations with reduced larval susceptibility to spinosad. Our findings show that modest standing variation can shape divergence at insecticide target sites within a highly constrained gene family and underscore the need to monitor interspecific variation during the deployment of nAChR-targeting insecticides.

evolutionary biology↗

CB2R-induced differentiation epigenetically restrains cancer plasticity enabling adaptive therapy

Cellular plasticity enables cancer cells to escape therapy by adopting stem-like or alternate lineage states. Here, we identify a mechanism by which cannabinoid receptor 2 (CB2R) activation promotes irreversible lineage commitment in breast cancer. Using patient-derived and murine organoids, we show that brief, low-dose exposure to CB2R agonists--either phytogenic or synthetic--induces a basal-to-luminal transition, accompanied by reduced self-renewal, invasiveness, and tumor-initiating potential. These changes are retained under conditions that normally promote dedifferentiation, including fibroblast co-culture, immune pressure, and mechanical shear stress. Mechanistically, CB2R engagement initiates a transient chromatin remodeling program, marked by early expression of pluripotency-associated genes followed by silencing and differentiation commitment. This epigenetically stabilized state renders tumor cells more responsive to tamoxifen and limits the emergence of resistant clones. Our findings uncover a previously unrecognized role for CB2R in modulating cancer cell identity and suggest new opportunities to constrain tumor plasticity by directing differentiation through a drug-responsive pathway.

cancer biology↗

The gut microbiome of the Passalid beetle has high cellulolytic potential and constitutes an unrecognized system for production of greenhouse gasses in neotropical forests

Herbivores, and their associated microbiomes, play a major role in the global carbon cycle. Recalcitrant cellulose molecules are broken down by microorganisms that colonize the herbivore gut, releasing greenhouse gasses in the process. Passalid beetles are tropical herbivores that feed only on decaying wood and present subsocial behavior that may lead to the acquisition and sharing of microbial symbionts for efficient biomass and energy production. We collected five groups of Veturius sp. Passalid beetles from different logs in the Costa Rican rainforest and analyzed the microbial communities of larval and adult guts, as well as the substrate material in which they resided (partially chewed wood material mixed with feces that covers their tunnels). Adults, larvae, and substrate harbor different microbial communities, with substrate showing the highest diversity and richness, and larvae gut comprising a high abundance of methanogenic archaea. Reconstructed metagenome assembled genomes (MAGs) revealed that larvae and adults are enriched in genomes encoding a myriad of glycosyl hydrolases. Methanogenic taxa were enriched within the larval MAGs and bins, suggesting that larval microbiota participate in the final steps of cellulose decomposition in the system. Finally, we assessed methane production rates by Passalid beetles and compared them with termites. Our results show that Passalid larvae and adults produce methane at rates comparable to termites. Passalid and other beetle larvae are potentially underappreciated contributors to the carbon cycle and the biotic production of the greenhouse gas methane.

microbiology↗

Light and sex modify Snord116 genotype effects on metabolism, behavior, and imprinted gene networks following circadian entrainment

Mammals utilize imprinted and X-linked epigenetic mechanisms in development, metabolism, and behavior. Imprinted genes, including Prader-Willi syndrome Snord116 noncoding RNAs, are implicated in the regulation of sleep and circadian rhythms through poorly understood mechanisms. Utilizing mouse models of Snord116 deficiency and overexpression, we performed an integrated, sex-stratified analysis of free running behaviors, indirect calorimetry, and cortical transcriptomes following entrainment to a 22 hour light:dark T-cycle. We observed significant interactions of sex, entrainment, and Snord116 genotype in period length at baseline and after-effects of post-entrainment. Snord116 deletions effect on respiratory exchange ratio was light sensitive, with sex and entrainment effects dominant under total darkness. Snord116 genotype impacted both rhythmic and non-rhythmic cortical gene networks that integrated sex, light, and entrainment effects with genotype-phenotype correlations. A co-expressed gene network enriched for imprinted, Snord116-target, and Xist-proximal long noncoding RNAs was identified as a light-sensitive regulatory hub of sexual dimorphic responses to a dynamic environment.

animal behavior and cognition↗

High-resolution HIV-1 m6A epitranscriptome reveals splicing-dependent methylation clusters and unique 2-LTR transcript modifications

The N6-methyladenosine (m6A) modification of HIV-1 has been widely studied but the number and precise positions of the m6A sites remain unclear due to the lack of precision of detection methods. Using the latest Nanopore chemistry and direct m6A base-calling, we identified 18 m6A: 14 at the 3 end and 4 in central regions of the genome. Our data reveal differential methylation of these positions between splicing isoforms. Eleven of these sites are clustered in two short segments with peak-shaped methylation profiles. Single-molecule analysis revealed that a very small number of transcripts were unmethylated in both clusters. We also identified a [~]732 nt RNA species resulting from the transcription of non-integrated viral DNA circles closed by two long terminal repeats. These transcripts started in the first LTR, terminated at the polyA site of the second LTR and harbored six m6A sites. Five of these sites were present in other transcripts and, remarkably, had the highest methylation rates. The sixth site was methylated only in this transcript, suggesting a role for this RNA in HIV-1 infection. These findings reveal a new landscape of HIV m6A transcriptome modifications and pave the way for studies deciphering their role in the viral life cycle.

bioinformatics↗

Modelling forest dynamics using integral projection models (IPMs) and repeat LiDAR

O_LIEstimating the life histories and population dynamics of trees is important for predicting how forests respond to climate change and disturbances. To do so, ecologist often use stage-structured population models, which explicitly account for individual heterogeneity. However, the data needed to parameterise these models is typically constrained by time- and labour-intensive field methods. C_LIO_LIDespite growing access to remote sensing imagery that could potentially satiate these data-hungry population models, these data remain underutilised in population ecology. Here, we demonstrate a pipeline that integrates repeat-LiDAR data with a type of structured demographic model (Integral Projection Model, IPM) to examine forest-wide dynamics in response to environmental drivers. Using Australias Great Western Woodlands as a case study, we model the survival and growth of [~]40,000 eucalypt trees over a decade to estimate their life expectancy and characterise multiple stages of growth (height growth and crown expansion). C_LIO_LIOur results indicate distinct responses of small trees (where vital rates are predicted by height) and large trees (which invested predominantly in expanding crown area) to proxies for competition and soil moisture (local canopy density and topographic wetness index, respectively). Parameterising structured population models using LiDAR data therefore offers a step-change in perspective towards more ecologically meaningful forest dynamics. C_LIO_LITo broaden the application of this pipeline, we highlight three priorities: (1) Application to more complex systems such as mixed species stands and dense, multilayered canopies); (2) Incorporating complete life histories, including reproduction and early life stages; and (3) Using in long-term or comparative studies through improved availability of high-quality, comparable, repeat-LiDAR surveys. By combining remote sensing data with detailed insights from field- based studies, this pipeline represents a scalable framework for guiding forest management and conservation decisions. C_LI

ecology↗

Design and Validation of a Wearable Imaging System for Automated Wound Monitoring in Porcine Model

Effective wound monitoring has the potential to guide treatment regiments and improve healing outcomes, yet current clinical assessment methods remain largely subjective and labor-intensive. To address this challenge, we present a high-resolution wearable imaging system designed for continuous wound monitoring. The system integrates a 64 MP camera with a plano-convex lens housed in an enclosure measuring 73 mm in diameter and 36.1 mm in height, and features a custom printed circuit board (PCB) for programmable LED illumination. The 3D-printed device enclosure is designed to accommodate a silicone bioelectronic device and can be securely attached using a commercially available ostomy skin barrier. In porcine model validation studies, the system successfully captured daily wound progression over periods up to 7 days. The captured images were wirelessly transmitted to a processing unit where DeepMapper, a machine learning algorithm, processed z-stacked images and performed multi-level feature extraction to predict wound healing stages and indicate potential complications such as infection. This imaging system enables automated analysis of wound progression and supports the development of smart wound care platforms for personalized treatment strategies. The integrated design approach demonstrates the feasibility of creating compact, high-resolution imaging systems suitable for clinical wound monitoring applications.

bioengineering↗

Wireless bioelectronic device for wound healing

Wireless bioelectronic actuators have been developed to deliver targeted treatments over multiple days while continuously monitoring delivery, thereby improving wound healing. Specifically, these devices can deliver charged biomolecules such as fluoxetine cations (Flx+) and electric field (EF) in freely moving pigs. Treatments can be controlled and monitored in real time via WiFi, with options for both user-specified delivery rates and durations, as well as automated closed-loop (CL) control. The devices are engineered to handle various failure scenarios that may arise in dynamic, real-world experiments--such as communication or power interruptions--ensuring that valuable experimental data is collected with minimal disruption. The ion pump features eight drug reservoirs and channels arranged around a 20 mm diameter-wound, with a central ground electrode (0 V). When voltages above 0 V are applied to the outer channels, currents flow from the reservoirs and channels into the wound, delivering Flx+ and/or EF depending on the reservoir solution. The device records applied voltages and currents locally to a microSD card at a high sampling rate, while simultaneously transmitting real-time measurements via a local WiFi network to a wound healing algorithm running on a nearby laptop. CL control of current/delivery rate is performed by an onboard microcontroller unit (MCU) and current-source microchips, based on instructions received from the wound healing algorithm. A graphical user interface (GUI) provides intuitive user control and real-time data visualization, with support for multiple devices. In vivo studies over seven days showed that Flx+-treated wounds had a 20% lower M1/M2 macrophage ratio and 41.67% greater re-epithelialization compared to controls (standard-of-care), demonstrating the actuators potential to enhance wound healing.

biochemistry↗

Variation in precipitation drives differences in interactions and short-term transient instability between grassland functional groups: a stage-structured community approach

Climate change is expected to increase the frequency and severity of precipitation extremes, causing droughts and flooding. Consequently, grassland communities are forecasted to become increasingly unstable. To predict grassland responses, we need empirical information together with models that reliably extrapolate community dynamics from those observations. However, such prediction is challenging because community models typically simulate long-term (asymptotic) performance, and thus potentially neglect their short-term (transient) performance. Here, we use data from a precipitation experiment performed over eight years to model both short- and long-term responses of three functional groups - grasses, legumes, and non-leguminous forbs - to precipitation extremes. We use multi-functional-group Integral Projection Models and pseudospectral theory, to track grassland community dynamics. We show that the percentage-cover-stage-structure of functional groups shapes their transient instability, and that inter-functional-group interactions are competitive under increased precipitation but facilitative under decreased precipitation. IPMs and pseudospectra enable forecasting of how functional-group-stage-structure drives responses to climatic extremes.

ecology↗

The malaria parasite Plasmodium falciparum prepares for life within the host cell by activating a complex gene network

The intraerythrocytic developmental cycle (IDC) during which the malaria parasite Plasmodium falciparum multiplies asexually initiates when parasite stages called merozoites are released into the bloodstream, following schizont rupture, and invade the host red blood cells. The merozoite is the less studied stage of the IDC, despite its importance for the establishment of infection. Here we uncovered the gene expression profile of the free merozoites, by doing single cell RNA-sequencing (scRNA-seq). We found that free merozoites express invasion and motility genes that are actively transcribed in schizonts and after merozoite egress. P. falciparum parasites undergo at least one IDC before sexually-committing and transforming into the sexual stages, the gametocytes, which are then transmitted to the mosquito vector. Our scRNA-seq dataset identified two early states of sexual commitment, preceding expression of the gametocytogenesis master regulator ap2-g. We hypothesize that early committed parasites are primed for sexual differentiation and definitive engagement into gametocytogenesis occurs when ap2-g is expressed. Our results contribute to a better understanding of how the parasite prepares for invasion and how it exits asexual growth and enters sexual differentiation. These transitions ensure survival of the parasite in the human host and transmission to the mosquito vector so the gene markers identified here can be therapeutically explored in the future to block malaria invasion or transmission. ImportanceThe malaria parasite Plasmodium falciparum, which causes the death of over half a million people each year, has a complex life cycle but symptoms only arise when the parasite invades and develops inside the host red blood cells. The merozoites are the parasite stages responsible for invading the human red blood cells and they are one of the few extracellular stages of the lifecycle, even if briefly. However, their gene expression profile had yet to be determined. Here we used single cell RNA-sequencing to establish the gene expression program of free merozoites. We also identified for the first time the gene signature of sexually-committed and sexual merozoites that will later give raise to gametocytes and ensure parasite transmission.

genomics↗

A double hit affecting the IKZF1-IKZF2 tandem in immune cells of schizophrenic patients regulate specific symptoms

Schizophrenia is a complex multifactorial disorder and increasing evidence suggests the involvement of immune dysregulations in its pathogenesis. We observed that IKZF1 and IKZF2, classic immune-related transcription factors (TFs), were both downregulated in patients peripheral blood mononuclear cells (PBMCs) but not in their brain. We generated a new mutant mouse model with a reduction in Ikzf1 and Ikzf2 to study the impact of those changes. Such mice developed deficits in the three dimensions (positive-negative-cognitive) of schizophrenic-like phenotypes associated with alterations in structural synaptic plasticity. We then studied the secretomes of cultured PBMCs obtained from human patients and identified potentially secreted molecules, which depended on IKZF1 and IKZF2 levels, and that in turn have an impact on neural synchrony, structural synaptic plasticity and schizophrenic-like symptoms in in vivo and in vitro models. Our results point out that IKZF1-IKZF2-dependent immune signals negatively impact on essential neural circuits involved in schizophrenia.

neuroscience↗

Heterologous Prime-Boost with Immunologically Orthogonal Protein Nanoparticles for Peptide Immunofocusing

Protein nanoparticles are effective platforms for antigen presentation and targeting effector immune cells in vaccine development. Encapsulins are a class of protein-based microbial nanocompartments that self-assemble into icosahedral structures with external diameters ranging from 24 to 42 nm. Encapsulins from Mxyococcus xanthus were designed to package bacterial RNA when produced in E. coli and were shown to have immunogenic and self-adjuvanting properties enhanced by this RNA. We genetically incorporated a 20-mer peptide derived from a mutant strain of the SARS-CoV-2 receptor binding domain (RBD) into the encapsulin protomeric coat protein for presentation on the exterior surface of the particle. This immunogen elicited conformationally-relevant humoral responses to the SARS-CoV-2 RBD. Immunological recognition was enhanced when the same peptide was presented in a heterologous prime/boost vaccination strategy using the engineered encapsulin and a previously reported variant of the PP7 virus-like particle, leading to the development of a selective antibody response against a SARS-CoV-2 RBD point mutant. While generating epitope-focused antibody responses is an interplay between inherent vaccine properties and B/T cells, here we demonstrate the use of orthogonal nanoparticles to fine-tune the control of epitope focusing. Table of Contents graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=121 SRC="FIGDIR/small/581861v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@53407corg.highwire.dtl.DTLVardef@1ae8d0eorg.highwire.dtl.DTLVardef@aec021org.highwire.dtl.DTLVardef@c7e0ef_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

Signal Transduction in an Enzymatic Photoreceptor Revealed by Cryo-Electron Microscopy

Phytochromes are essential photoreceptor proteins in plants with homologs in bacteria and fungi that regulate a variety of important environmental responses. They display a reversible photocycle between two distinct states, the red-light absorbing Pr and the far-red light absorbing Pfr, each with its own structure. The reversible Pr to Pfr photoconversion requires covalently bound bilin chromophore and regulates the activity of a C-terminal enzymatic domain, which is usually a histidine kinase (HK). In plants, phytochromes translocate to nucleus where the C-terminal effector domain interacts with protein interaction factors (PIFs) to induce gene expression. In bacteria, the HK phosphorylates a response-regulator (RR) protein triggering downstream gene expression through a two-component signaling pathway. Although plant and bacterial phytochromes share similar structural composition, they have contrasting activity in the presence of light with most BphPs being active in the dark. The molecular mechanism that explains bacterial and plant phytochrome signaling has not been well understood due to limited structures of full-length phytochromes with enzymatic domain resolved at or near atomic resolution in both Pr and Pfr states. Here, we report the first Cryo-EM structures of a wild-type bacterial phytochrome with a HK enzymatic domain, determined in both Pr and Pfr states, between 3.75 and 4.13 [A] resolution, respectively. Furthermore, we capture a distinct Pr/Pfr heterodimer of the same protein as potential signal transduction intermediate at 3.75 [A] resolution. Our three Cryo-EM structures of the distinct signaling states of BphPs are further reinforced by Cryo-EM structures of the truncated PCM of the same protein determined for the Pr/Pfr heterodimer as well as Pfr state. These structures provide insight into the different light-signaling mechanisms that could explain how bacteria and plants see the light.

molecular biology↗