Biology subjects

Schuler, H.

Publications and source records attributed to Schuler, H..

At least 19 recordsLinked to original sources

bioRxiv2025

Sex-specific exploration accounts for differences in valence learning in male and female mice

Valence, the quality by which something is perceived as good or bad, appetitive or aversive, is a fundamental building block of emotional experience and a primary driver of adaptive behavior. Pavlovian fear and reward learning paradigms are widely used in preclinical research to probe mechanisms of valence learning but with limited consideration of sex as a biological variable despite known sex differences in neuropsychiatric disorders associated with impaired valence. Here, we compare appetitive-only, aversive-only and mixed-valence cue-outcome Pavlovian conditioning paradigms in male and female mice to dissociate effects of context, valence and salience in a sex-specific manner. Using a data-driven approach to identify behaviors indicative of valence learning in an unbiased manner, we compare task performance between paradigms in male and female mice. We show that while male and female mice acquire appetitive and aversive associations in both single- and mixed-valence paradigms, sex differences emerge in single-valence paradigms. Ultimately, we show that these apparent sex differences in valence learning are driven by non-specific baseline differences in exploratory behavior. Males explore more at baseline, altering their trajectory of cue-reward association acquisition whereas females explore less at baseline, increasing shock facilitated freezing in aversive-only contexts, masking cue discrimination. Overall, our findings illustrate how task design differentially impacts behavioral expression in male and female mice and demonstrate that mixed-valence paradigms afford a more accurate assessment of valence learning in both sexes.

neuroscience↗

bioRxiv2025

Rapid evolution of Wolbachia in cherry fruit flies

Wolbachia is a widespread bacterial endosymbiont in arthropods known for its ability to manipulate host reproduction through cytoplasmic incompatibility (CI), thereby promoting its spread. However, Wolbachia harbors an array of mobile genetic elements (MGEs) that can rapidly alter its genomic structure and content, including the cif loci responsible for CI. At the phylogenetic scale, Wolbachia genomes are shown to be dynamic with MGEs causing widespread genome rearrangements. But on ecological time scales, the evolution of Wolbachia remains largely unknown. In this study, we leverage the natural history of Wolbachia in cherry-infesting fruit flies in the Rhagoletis cingulata sibling species group. Members of this species group share a common Wolbachia strain, wCin2, and their divergence spans from 1,000 to 150,000 years ago. We utilized Nanopore sequencing to characterize wCin2 Wolbachia genome divergence across recent to more distant evolutionary timescales. We report rapid evolution of population-level differences in gene content (including cif loci and MGEs) and genome structure, with differentiation increasing with time since host divergence. Notably, structural variants were the first to appear both between and among wCin2 populations. Our results also indicated that the CI phenotype previously attributed to a distinct Wolbachia strain, wCin3, that coinfects flies in the southwestern USA and Mexico, may instead be caused by cif genes that were horizontally transferred into wCin2. Finally, we discovered a novel Wolbachia strain, wInd, which appears to have been recently horizontally acquired by cherry fly populations in the Pacific Northwest of the USA. Our findings underscore the fluidity and rapid genome evolution of Wolbachia, with significant implications for cif gene dynamics and potential impacts on host evolutionary trajectories. Author SummaryIn our study, we explore the genome evolution of Wolbachia, a common bacterial endosymbiont found in many insects that can manipulate its hosts reproduction to help it spread more effectively. Wolbachias genome is shown to change dramatically over long evolutionary periods due to mobile genetic elements. However, little is known about how Wolbachias genome evolves over shorter ecological timeframes. Here, we utilized cherry-infesting fruit flies and their Wolbachia strain known as wCin2, both of which have spread across North America for the past 150,000 years. Using long-read sequencing, we assembled multiple wCin2 Wolbachia genomes from across North America and made comparisons between wCin2 Wolbachia genomes that have recently diverged and comparisons to those that have been isolated for over 150,000 years. Our findings reveal that Wolbachias genome can evolve swiftly, with differences in gene content and most prominently genome structure arising between recently separated Wolbachia populations and increasing as populations diverge over time. Interestingly, these changes also impact the key genes responsible for Wolbachias reproductive manipulation. This research highlights the rapid and dynamic nature of Wolbachia genome evolution across different evolutionary timescales and offers insights into endosymbiont-host relationships as well as their potential impact on host speciation and vector control.

evolutionary biology↗

bioRxiv2025

Single-cell sequencing reveals psilocybin induces sustained cell-type specific plasticity in mouse medial prefrontal cortex

Despite enormous interest in psychedelics for psychiatric interventions, potential underlying biological mechanisms remain unclear. Here, we confirm that a single dose of psilocybin increases synaptic transmission in mouse medial prefrontal cortex. Using scRNA-sequencing, we identify cell-type specific mechanisms of sustained neuroplastic effects. We show that, 24h post-psilocybin, expression of plasticity-related genes is increased in excitatory neurons and that transcription in a type of deep layer near projecting neuron, L5/6 NP, is robustly altered. Analyzing receptor expression patterns reveals that this cell-type specificity does not align with 5-HT2A expression but aligns with 5-HT2C expression patterns. Further, multivariate analyses identify psilocybin-induced gene expression patterns in L5/6 NP neurons predict 5-HT2C, but not 5-HT2A, transcript levels. Pharmacologic manipulation with a 5-HT2C antagonist attenuates the post-acute sustained effect of psilocybin on synaptic transmission, highlighting 5-HT2C signaling and L5/6 NP neurons as key mediators of psychedelic drug actions sustained neuroplastic effects in mPFC.

neuroscience↗

bioRxiv2024

Deciphering the patterns and timing of diversification of the genus Melanastera (Hemiptera: Psylloidea: Liviidae) in the Neotropics

Even after decades of research on diversification in the Neotropics, our understanding of the evolutionary processes shaping Neotropical clades is still incomplete. In the current study, we used different divergence times and likelihood-based methods to investigate the influence of biogeography and host associations on the diversification of the most species-rich Neotropical psyllid genus Melanastera (Liviidae) using molecular phylogenetic data from seven gene fragments (four mitochondrial and three nuclear). The putatively monophyletic group of Neotropical Melanastera species has an estimated crown node age of 20.2 Ma (ML, CI 20.2-30.6) or 23.2 Ma (BI, 95% HPD 16.6-32.6), with diversification occurring mainly in the Upper Miocene, although some species groups diversified in the Pliocene and Pleistocene. Biogeographic analysis suggests that the Neotropical Melanastera originated from the Pacific region of South and Central America. We detected a shift in diversification rates that likely occurred either at the time of origin of Melanastera or during the main colonisation of the Atlantic and Amazon Forests, followed by a subsequent slowdown in speciation rates. State-dependent speciation and extinction models revealed a significant relationship between this diversification shift and the shift of Melanastera to the plant families Melastomataceae and Annonaceae, reflecting the impact of host switching on speciation rates in this group. This period also coincides with several independent dispersal events from the Atlantic and Amazon Forests to other parts of the Neotropics. Taken together, the results of the current study suggest that diversification of Melanastera was facilitated by major shifts to new host families, which may have promoted the dispersal of Melanastera into new adaptive zones with subsequent processes of local speciation.

evolutionary biology↗

bioRxiv2024

Data-driven analysis identifies female-specific social modulation deficit after chronic social defeat stress

BackgroundChronic social defeat stress is a widely used depression model in male mice. Several proposed adaptations extend this model to females with variable, often marginal effects. We examine the if widely used male-defined metrics of stress are suboptimal in females and reveal sex-specific adaptations. MethodsUsing a data-driven method we comprehensively classified social interaction behavior in 761 male and female mice after chronic social witness/defeat stress, examining social modulation of behavioral frequencies and associations with conventional metrics (i.e., social interaction (SI) ratio). ResultsSocial stress induces distinct behavioral adaptation patterns in males and females. SI ratio leads to underpowered analyses in females with limited utility to differentiate susceptibility/resilience. Data-driven analyses reveal failure of social adaptation in stressed female mice that is captured in attenuated velocity change from no target to target tests ({Delta}Velocity) and validate this in three female social stress models. Combining SI ratio and {Delta}Velocity optimally differentiates susceptibility/ resilience in females and this metric reveals resilient-specific adaptation in a resilience-associated neural circuit in female mice. ConclusionsWe demonstrate that psychological or physical social defeat stress induces similar deficits in females that is qualitatively distinct from male deficits and inadequately sampled by male-defined metrics. We identify modulation of locomotion as a robust and easily implementable metric for rigorous research in female mice. Overall, our findings highlight the need to critically evaluate sex differences in behavior and implement sex-based considerations in preclinical model design.

neuroscience↗

bioRxiv2024

Regulation of ADP-ribosyltransferase activity by ART domain dimerization in PARP15

PARP15 is a mono-ADP-ribosyltransferase with unknown functions. Its evolutionary relationship with PARP14 suggests roles in antiviral defense; its ability to modify RNA and localization to stress granules point to functions in the regulation of translation. PARP15 also modifies itself and other proteins using its ADP-ribosyltransferase (ART) domain and contains two macrodomains predicted to bind ADP-ribosyl on targets. We used biochemical and biophysical analysis to study how the ADP-ribosyltransferase activity of PARP15 is regulated. Here we show that the catalytic domain of PARP15 dimerizes with mid-nanomolar affinity, forming the same dimer interface in solution that had already been captured by X-ray crystallography of the domain. Furthermore, we show that the formation of dimers is a prerequisite for catalytic activity and that monomeric mutant variants of the domain were catalytically inactive. Our findings suggest a regulatory mechanism by which dimerization is linked to either target engagement or placement of a catalytic residue, rather than NAD+ co-substrate binding, and by which the two protomers of the dimer operate independent of one another. Together, our results uncover a novel mechanism of regulation in a PARP family enzyme, which might inspire new avenues of pharmacological intervention.

biochemistry↗

bioRxiv2024

Disentangling plant response to biotic and abiotic stress using HIVE, a novel tool to perform unpaired multi-omics integration

All organisms are subjected to multiple stresses usually occurring at the same time, requiring the activation of the appropriate signalling pathways to respond to all or by prioritizing the response to one stress factor. Plants, as sessile organisms, are particularly impacted by the constantly changing environment that is often unfavourable or even hostile. Because of the experimental complexity of studying the response of one organism to multiple stressors simultaneously, usually experiments are conducted considering one individual stress factor at the time. An alternative consists in performing in silico integration of those data on single stress response. Currently used methods to integrate unpaired experiments consist of performing meta-analysis or finding differentially expressed genes for each condition separately and then selecting the commonly regulated ones. Although these approaches allowed to find valuable results, they mainly identify specific signatures in response to one stress and very few signature responding to multiple stresses and lack those modulated differently in each condition. For this purpose, we developed HIVE (Horizontal Integration analysis using Variational AutoEncoders) to integrate multiple single-stress transcriptomics datasets composed of unpaired experiments. Briefly, we coupled a variational autoencoder, that alleviates batch effects, with a random forest regression and the SHAP explainer to select relevant genes modulated specifically in response to one or multiple stresses. We illustrate the functionality of HIVE to study the transcriptional changes of several different plants namely Arabidopsis thaliana, rice, maize, wheat, grapevine and peanut by collecting publicly available experiments on single stress, either biotic and/or abiotic, and jointly analyse them. HIVE performed better than the differential expression analysis, meta-analysis and the state-of-the-art tool for horizontal integration allowing to identify novel promising candidates responsible for triggering effective defence responses to multiple stresses.

plant biology↗

bioRxiv2023

Definition of the effector landscape across 13 Phytoplasma proteomes with LEAPH and EffectorComb

BackgroundCrop pathogens are a major threat to plants health, reducing the yield and quality of agricultural production. Among them, the Candidatus Phytoplasma genus, a group of fastidious phloem-restricted bacteria, can parasite a wide variety of both ornamental and agro-economically important plants. Several aspects of the interaction with the plant host are still unclear but it was discovered that phytoplasmas secrete certain proteins (effectors) responsible for the symptoms associated with the disease. Identifying and characterizing these proteins is of prime importance for globally improving plant health in an environmentally friendly context. ResultsWe challenged the identification of phytoplasmas effectors by developing LEAPH, a novel machine-learning ensemble predictor for phytoplasmas pathogenicity proteins. The prediction core is composed of four models: Random Forest, XGBoost, Gaussian, and Multinomial Naive Bayes. The consensus prediction is achieved by a novel consensus prediction score. LEAPH was trained on 479 proteins from 53 phytoplasmas species, described by 30 features accounting for the biological complexity of these protein sequences. LEAPH achieved 97.49% accuracy, 95.26% precision, and 98.37% recall, ensuring a low false-positive rate and outperforming available state-of-the-art methods for putative effector prediction. The application of LEAPH to 13 phytoplasma proteomes yields a comprehensive landscape of 2089 putative pathogenicity proteins. We identified three classes of these proteins according to different secretion models: "classical", presenting a signal peptide, "classically-like" and "non-classical", lacking the canonical secretion signal. Importantly, LEAPH was able to identify 15 out of 17 known experimentally validated effectors belonging to the three classes. Furthermore, to help the selection of novel candidates for biological validation, we applied the Self-Organizing Maps algorithm and developed a shiny app called EffectorComb. Both tools would be a valuable resource to improve our understanding of effectors in plant-phytoplasmas interactions. ConclusionsLEAPH and EffectorComb app can be used to boost the characterization of putative effectors at both computational and experimental levels and can be employed in other phytopathological models. Both tools are available at https://github.com/Plant-Net/LEAPH-EffectorComb.git.

bioinformatics↗

bioRxiv2023

Identification and characterization of specific motifs in effector proteins of plant parasites using MOnSTER.

Plant pathogens cause billions of dollars of crop loss every year and are a major threat to global food security. Identifying and characterizing pathogens effectors is crucial towards their improved control. Because of their poor sequence conservation, effector identification in protein sequences predicted from genomes is challenging and current methods generate too many candidates without indication for prioritizing further experimental studies. In most phyla, effectors contain specific sequence motifs which influence their localization and targets in the plant. Although bacterial, fungal and oomycetes effectors have been studied extensively and conserved characteristic motifs have been identified, research on plant-parasitic nematode effectors (PPN) identified some enriched degenerate motifs in only one species so far. The different lifestyles of PPNs might reflect effectors with different functions according to the nematodes specific needs, thus presenting a high variety of characteristic motifs. To circumvent these limitations, we have developed MOnSTER a novel tool that identifies clusters of motifs of protein sequences (CLUMPs). MOnSTER can be fed with motifs identified by de novo tools or from databases such as Pfam and InterProScan. The advantage of MOnSTER is the reduction of motif redundancy by clustering them and associating a score. This score encompasses the physicochemical properties of AAs and the motif occurrences. We built up our method to identify discriminant CLUMPs in candidate parasitism proteins of plant-pathogenic oomycetes. We showed the reliability of MOnSTER by identifying five CLUMPs that correspond to the known motifs: RxLR, -dEER and LxLFLAK-HVLVxxP. Consequently, we applied MOnSTER on PPN candidate parasitism proteins and identified peculiar motifs in their sequences. We identified six CLUMPs in about 60% of the known nematode candidate parasitism proteins. Furthermore, we found that specific co-occurrences of at least two CLUMPs are present in PPN candidate parasitism protein sequences bearing protein domains important for invasion and pathogenicity. The potentiality of this tool goes beyond the candidate parasitism proteins and can be used to easily cluster motifs and calculate the CLUMP-score on any set of protein sequences. Authors summaryPopulation growth, environmental degradation and climate change are already bringing harm to human communities and the natural world that needs to be addressed rapidly. Ensuring food security for a population that will exceed 9 billion people by 2050 while preserving the environment and biodiversity is a major challenge. Agricultural pathogens, to cause the infection, secrete effector proteins that promote colonization of the host plant. Identifying and characterizing pathogens effectors is crucial towards understanding how they manipulate the plant and better combat them. Because of their poor sequence conservation, effector identification in protein sequences predicted from genomes is challenging and current methods generate too many candidates without indication for prioritizing further experimental studies. To address these challenges, we have developed a novel tool called MOnSTER, that identifies and score clusters of motifs of protein sequences (CLUMPs). MOnSTER is an easy to use tool that can be included in any pipeline needing motif calling and will be of great use to accelerate both computational and experimental studies relating to protein motif discovery. Altogether our findings provide improvements in the understanding of the mechanisms set up by the pathogens to infect the plant and can elucidate important signatures to block the development of plant-pathogen interactions and allow to engineer of durable disease resistance.

bioinformatics↗

bioRxiv2023

PARP14 is a writer, reader and eraser of mono-ADP-ribosylation

PARP14/BAL2 is a large multidomain enzyme involved in signaling pathways with relevance to cancer, inflammation, and infection. Inhibition of its mono-ADP-ribosylating PARP homology domain and its three ADP-ribosyl binding macro domains has been regarded as a potential means of therapeutic intervention. Macrodomains-2 and -3 are known to stably bind to ADP-ribosylated target proteins; but the function of macrodomain-1 has remained some-what elusive. Here, we used biochemical assays of ADP-ribosylation levels to characterize PARP14 macrodomain-1 and the homologous macrodomain-1 of PARP9. Our results show that both macrodomains display an ADP-ribosyl glycohydrolase activity that is not directed toward specific protein side chains. PARP14 macrodomain-1 is unable to degrade poly(ADP-ribose), the enzymatic product of PARP1. The F926A mutation of PARP14 and the F244A mutation of PARP9 strongly reduced ADP-ribosyl glycohydrolase activity of the respective macrodomains, suggesting mech-anistic homology to the Mac1 domain of the SARS-CoV-2 Nsp3 protein. This study adds two new enzymes to the previously known six human ADP-ribosyl glycohydrolases. Our results have key implications for how PARP14 and PARP9 will be studied and how their functions will be understood.

biochemistry↗

bioRxiv2023

The bacterial community of the European spruce bark beetle in space and time

The European spruce bark beetle Ips typographus is a pest causing severe damages in forests dominated by the Norway spruce in Europe. Microorganisms play an essential role in the host species performance, including nutrition, fitness as well as in overcoming host defenses. Here, we performed high-throughput 16S rRNA metabarcoding of I. typographus across different populations in Europe, to assess its bacterial community. We investigated four postglacial refugial areas in Europe and focused specifically on a current bark beetle hot spot in the Dolomites where we compared populations with different epidemiological phases (outbreaking vs. non-outbreaking) and across different seasons (pre-overwintering vs. overwintering). Our results show that the bacterial community structure varied among populations from the refugial areas and between different regions within the Dolomites. We found a significant difference in the bacterial community between pre-overwintering and overwintering individuals, but we did not find differences between epidemic and endemic populations. The prevalence of the genus Erwinia which was present in every individual and Pseudoxanthomonas in almost every individual across all populations, suggests that these taxa form the core bacterial community of I. typographus. Furthermore, several additional bacterial taxa occurred in all populations, but with variable frequencies. This study highlights a complex interaction of I. typographus and various bacterial taxa across different regions and ecological phases of I. typographus populations and provides new insights into the role of microorganisms in the biology of this important pest species.

ecology↗

bioRxiv2023

Division of labour within psyllids: Metagenomics reveals an ancient dual endosymbiosis with metabolic complementarity in the genus Cacopsylla

Hemipteran insects are well-known for their ancient associations with beneficial bacterial endosymbionts, particularly nutritional symbionts providing the host with essential nutrients such as amino acids or vitamins lacking from the hosts diet. Thereby, these primary endosymbionts enable the exploitation of nutrient-poor food sources such as plant sap or vertebrate blood. In turn, the strictly host-associated lifestyle strongly impacts the genome evolution of the endosymbionts, resulting in small and degraded genomes. Over time, even the essential nutritional functions can be compromised, leading to the complementation or replacement of an ancient endosymbiont by another, more functionally versatile, bacterium. Herein, we provide evidence for a dual primary endosymbiosis in several psyllid species. Using metagenome sequencing, we produced the complete genome sequences of both the primary endosymbiont Candidatus Carsonella ruddii and an as yet uncharacterized Enterobacteriaceae bacterium from four species of the genus Cacopsylla. The latter represents a new psyllid-associated endosymbiont clade for which we propose the name Candidatus Psyllophila symbiotica. Fluorescent in situ hybridisation confirmed the co-localization of both endosymbionts in the bacteriome. The metabolic repertoire of Psyllophila is highly conserved across host species and complements the tryptophan biosynthesis pathway that is incomplete in the co-occurring Carsonella. Unlike co-primary endosymbionts in other insects, the genome of Psyllophila is almost as small as the one of Carsonella, indicating an ancient co-obligate endosymbiosis rather than a recent association to rescue a degrading primary endosymbiont. IMPORTANCEHeritable beneficial bacterial endosymbionts have been crucial for the evolutionary success of numerous insects, enabling the exploitation of nutritionally limited food sources such as vertebrate blood and plant sap. Herein, we describe a previously unknown dual endosymbiosis in the psyllid genus Cacospylla, consisting in the primary endosymbiont Candidatus Carsonella ruddii and a co-occurring Enterobacteriaceae bacterium for which we propose the name Candidatus Psyllophila symbiotica. Its localization within the bacteriome and its small genome size confirm that Psyllophila is a co-primary endosymbiont widespread within the genus Cacopsylla. Despite its highly eroded genome, Psyllophila complements the tryptophan biosynthesis pathway that is incomplete in the co-occurring Carsonella. Moreover, the genome of Psyllophila is almost as small as the one of Carsonella, indicating an ancient dual endosymbiosis rather than a recent acquisition of a new symbiont. Hence, our results shed light on the dynamic interactions of psyllids and their endosymbionts over evolutionary time.

evolutionary biology↗

bioRxiv2021

Effects of early life adversity on immediate early gene expression: systematic review and 3-level meta-analysis of rodent studies

Early-life adversity (ELA) causes long-lasting structural and functional changes to the brain, rendering affected individuals vulnerable to the development of psychopathologies later in life. Immediate-early genes (IEGs) provide a potential marker for the observed alterations, bridging the gap between activity-regulated transcription and long-lasting effects on brain structure and function. Several heterogeneous studies have used IEGs to identify differences in cellular activity after ELA; systematically investigating the literature is therefore crucial for comprehensive conclusions. Here, we performed a systematic review on 39 pre-clinical studies in rodents to study the effects of ELA on IEG expression. Females and IEGs other than cFos were investigated in only a handful of publications. We meta-analyzed publications investigating specifically cFos expression. ELA increased cFos expression after an acute stressor only if the animals (control and ELA) had experienced multiple negative life events. At rest, ELA increased cFos expression irrespective of other life events, suggesting that ELA creates a phenotype similar to naive, acutely stressed animals. We present a conceptual theoretical framework to interpret the unexpected results. Overall, ELA likely alters IEG expression across the brain, especially in interaction with other negative life events. The present review highlights current knowledge gaps and provides guidance to aid the design of future studies.

neuroscience↗

bioRxiv2021

Pipeline for 4D whole-brain microscopy preprocessing and analysis.

Acute stress leads to sequential activation of functional brain networks. The challenge is to get insight in whole brain activity at multiple scales, beyond the level of (networks of) nuclei. We developed a novel pre-processing and analytical pipeline to chart whole-brain immediate early genes expression - as proxy for cellular activity - after a single stressful foot-shock in 4 dimensions; that is, from functional networks up to 3D single-cell resolution, and over time. The pipeline is available as R-package. Most brain areas (96%) showed increased numbers of c-fos+ cells after foot-shock, yet hypothalamic areas stood out as being most active and prompt in their activation, followed by amygdalar, prefrontal, hippocampal and finally thalamic areas. At the cellular level, c-fos+ density clearly shifted over time across subareas, as illustrated for the basolateral amygdala. Moreover, some brain areas showed increased numbers of c-fos+ cells, while others dramatically increased c-fos intensity in just a subset of cells; this strategy changed after foot-shock in half of the brain areas. All single-cell data can be visualized for each of the 90 brain areas examined through our interactive web-portal.

neuroscience↗

bioRxiv2020

Stimulus-specific plasticity in human visual gamma-band activity and functional connectivity

Under natural conditions, the visual system often sees a given input repeatedly. This provides an opportunity to optimize processing of the repeated stimuli. Stimulus repetition has been shown to strongly modulate neuronal-gamma band synchronization, yet crucial questions remained open. Here we used magnetoencephalography in 30 human subjects and find that gamma decreases across ~10 repetitions and then increases across further repetitions, revealing plastic changes of the activated neuronal circuits. Crucially, changes induced by one stimulus did not affect responses to other stimuli, demonstrating stimulus specificity. Changes partially persisted when the inducing stimulus was repeated after 25 minutes of intervening stimuli. They were strongest in early visual cortex and increased interareal feedforward influences. Our results suggest that early visual cortex gamma synchronization enables adaptive neuronal processing of recurring stimuli. These and previously reported changes might be due to an interaction of oscillatory dynamics with established synaptic plasticity mechanisms.

neuroscience↗

bioRxiv2020

The genomics of trait combinations and their influence on adaptive divergence

Understanding rapid adaptation requires quantifying natural selection on traits and elucidating the genotype-phenotype relationship for those traits. However, recent studies have often failed to predict the direction of adaptive allelic variation in natural populations from laboratory studies. Here, we test for genomic signatures of genetic correlations to illustrate how multifarious, correlational selection may drive counterintuitive patterns of population divergence in the apple maggot fly, Rhagoletis pomonella (Diptera: Tephritidae). Apple-infesting populations with relatively early emerging adults have recently evolved from hawthorn-infesting populations consisting of relatively later emerging adults. Multiple studies have reported a paradoxical relationship between population differentiation and seasonal timing, as determined by the timing of diapause termination; alleles associated with late termination occur at higher frequencies in the earlier emerging apple-infesting populations compared to hawthorn-infesting populations. We present evidence that historical selection on diapause termination and another trait, initial diapause intensity, over geographic gradients generated genetic correlations between the traits in a direction antagonistic to contemporary selection on apple-infesting populations. Single nucleotide polymorphism in genomic regions of high linkage disequilibrium associated strongly with diapause termination and intensity, population divergence, geography, and evolutionary responses in laboratory selection experiments. These associations were consistent with geographically variable selection and with correlated evolutionary responses driving higher frequencies of late-associated alleles in the early emerging apple race. In contrast, loci associated only with diapause termination showed the expected pattern (more early-associated alleles in the apple race) in half of the population pairs. Our results suggest that selection on loci demonstrating antagonistic pleiotropy may often shape genomic footprints of rapid adaptation.

evolutionary biology↗

bioRxiv2018

System-wide Identification of Enzyme Substrates by Thermal Analysis (SIESTA)

Despite the immense importance of enzyme-substrate reactions, there is a lack of generic and unbiased tools for identifying and prioritizing substrate proteins which are modulated in the structural and functional levels through modification. Here we describe a high-throughput unbiased proteomic method called System-wide Identification and prioritization of Enzyme Substrates by Thermal Analysis (SIESTA). The approach assumes that enzymatic post-translational modification of substrate proteins might change their thermal stability. SIESTA successfully identifies several known and novel substrate candidates for selenoprotein thioredoxin reductase 1, protein kinase B (AKT1) and poly-(ADP-ribose) polymerase-10 systems in up to a depth of 7179 proteins. Wider application of SIESTA can enhance our understanding of the role of enzymes in homeostasis and disease, open new opportunities in investigating the effect of PTMs on signal transduction, and facilitate drug discovery.

biochemistry↗

bioRxiv2018

14-3-3 proteins activate Pseudomonas exotoxins-S and -T by chaperoning a hydrophobic surface

Pseudomonas are a common cause of hospital acquired infections that may be lethal. ADP-ribosyltransferase activities of Pseudomonas exotoxin-S and -T depend on 14-3-3 proteins inside the host cell. By binding in the 14-3-3 phosphopeptide binding groove, a hydrophobic C-terminal helix of ExoS and ExoT has been thought to be crucial for their activation. However, crystal structures of the 14-3-3{beta}:ExoS and -ExoT complexes presented here reveal an extensive novel binding interface that is sufficient for complex formation and toxin activation. We show that C-terminally truncated ExoS ADP-ribosyltransferase domain lacking the hydrophobic binding motif is active when co-expressed with 14-3-3. Moreover, swapping the hydrophobic C-terminus with a fragment from Vibrio Vis toxin creates a 14-3-3 independent toxin that ADP-ribosylates known ExoS targets. Finally, we show that 14-3-3 stabilizes ExoS against thermal aggregation. Together, this indicates that 14-3-3 proteins activate exotoxin ADP-ribosyltransferase domains by chaperoning their hydrophobic surfaces independently of the hydrophobic C-terminal segment.\n\nShort summaryCrystal structures of Pseudomonas exotoxins-S and -T identify a novel hydrophobic interface with 14-3-3 proteins, and we show that 14-3-3 activates these toxins independent of their phosphopeptide groove binding C-termini, by preventing their aggregation.

biochemistry↗