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Page, R.

Publications and source records attributed to Page, R..

15 recordsLinked to original sources

SHP2 binds directly to SOS1 to enable RAS activation

The protein-tyrosine phosphatase SHP2 (PTPN11) regulates growth factor- and cytokine-induced RAS/ERK MAP kinase (MAPK) pathway activation, and aberrant SHP2 function causes developmental disorders and cancer1-5. It is widely believed that the catalytic activity of SHP2 is essential for pathway activation1,4,6-8. This view has shaped our interpretation of how germline PTPN11 mutations cause Noonan Syndrome (NS) and NS with Multiple Lentigines (NS-ML)2,9 and how somatic mutations contribute to myeloproliferative neoplasms and solid tumors1. Here we identify a previously undetected, protein-tyrosine phosphatase (PTP) activity-independent mechanism that revises our understanding of how SHP2 promotes RAS/ERK activation. We find that certain mutations of the nucleophilic cysteine that abolish catalytic activity still promote RAS/ERK pathway activation in normal and neoplastic mammalian cells, zebrafish embryos, and mice. Structural studies show that the SHP2 PTP domain binds directly to the Son of Sevenless 1 (SOS1) Dbl homology (DH) domain. Proximity labeling and super-resolution microscopy demonstrate that SHP2/SOS1 interaction occurs in cells and facilitates SOS1 translocation to the plasma membrane to form clusters. Our results overturn decades of dogma on SHP2 regulation of the RAS/ERK pathway and provide new insights into the mechanism of action of disease-associated PTPN11 mutations.

Cancer Biology↗

Tracing the invisible: Quantifying mirroring and embodied attunement in dyadic and triadic Dance Movement Therapy

BackgroundMirroring is a foundational method used in Dance Movement Therapy (DMT), assumed to foster empathy and therapeutic attunement, yet its embodied dynamics remain insufficiently studied. In this paper, we provide the first quantitative exploration of client-therapist mirroring across dyadic and triadic formats, examining how synchrony unfolded during a structured mirroring exercise in which participants alternated between leading and following roles. MethodologyUsing optical motion capture and time-series modelling, we quantified movement coordination in dyadic (female client-therapist; male client-therapist) and triadic (therapist with both clients) interactions. ResultsIn dyadic tasks, the female client-therapist interaction was marked by tight temporal alignment, significant synchrony, robust predictive accuracy, and clear client-to-therapist influence, consistent with kinaesthetic empathy and affect-sensitive entrainment. By contrast, the male client-therapist dyad exhibited weaker and more delayed temporal coupling, alongside reduced phase synchronisation and fewer directional dependencies, despite comparable levels of interpersonal proximity. In the triadic task, temporal entrainment attenuated: therapist movement had few matching qualities to clients movement, yet recurrent synchrony with both clients persisted, suggesting a strategic shift from fine-grained entrainment to stable postural scaffolding under divided attention. DiscussionThese findings demonstrated that mirroring is not a uniform technique, but a family of embodied coordination modes flexibly recruited according to relational context and client expressivity. They align with theories of embodied simulation and affect attunement, implicating rapid motor resonance in dyadic entrainment and interoceptive-affective scaffolding in triadic stability. Clinically, the results underscore the need for training in flexible embodied strategies, split attention, and equitable allocation of attunement in group work. More broadly, they open a translational agenda linking kinematic synchrony to neural, interoceptive, and autonomic mechanisms, positioning mirroring as both an experiential hallmark and a measurable mechanism of change in embodied psychotherapy.

neuroscience↗

Defying expectations: sex-biased telomere dynamics and extended lifespan in the tropical bat species, Molossus molossus.

Telomeres are key biomarkers of cellular ageing, yet their dynamics remain poorly studied in tropical and short-lived bat species. Here, we present the first investigation of telomere length across age in Molossus molossus, a tropical bat historically categorised as the shortest-lived bat on record. Through a multi-year mark-recapture study in Gamboa, Panama, we sampled 492 individuals (n = 317 females, 175 males) and documented a female M. molossus surviving to at least 13 years of age, more than doubling the previously reported maximum lifespan of 5.6 years. Across the population, relative telomere length (rTL) showed no overall significant decline with age. No evidence was found for sex-specific rates of telomere attrition. Rather these results suggest that males and females follow parallel age-related telomere trajectories, with any sex differences primarily reflecting differences in mean telomere length rather than ageing dynamics. Overall, the findings here challenge previous assumptions about the lifespan and ageing biology of M. molossus. They demonstrate that telomere maintenance is not limited to temperate bats, show that sex differences in telomere biology are subtle and species-specific, and reinforce the value of long-term field studies for understanding ageing processes in the wild.

molecular biology↗

Mechanistic and evolutionary insights into a family of aminoacyl-tRNA deacylases that protects against canavanine toxicity

Aminoacyl-tRNA deacylases safeguard the accurate translation of the genetic code by hydrolyzing incorrectly synthesized aminoacyl-tRNAs. Canavanyl-tRNA deacylase (CtdA) was recently shown to protect cells against the toxicity of the non-proteinogenic amino acid canavanine, which is synthesized and accumulated by various plants. In most organisms, canavanine is ligated to tRNAArg, causing translation of arginine codons with canavanine. CtdA prevents canavanine toxicity by hydrolyzing canavanyl-tRNAArg. Here, we investigated the function, structure, substrate specificity, phylogenetic distribution, and evolution of CtdA. We show that CtdA is essential to prevent canavanine cytotoxicity in Salmonella enterica, and its heterologous expression can also rescue Escherichia coli. By determining the structure of CtdA, we identified its putative binding pocket and residues that modulate enzymatic activity and specificity. We also found that CtdA displays a relaxed specificity for the aminoacyl moiety substrate as it hydrolyzes arginyl-tRNAArg. Finally, we showed that despite their structural homology, CtdA and the aminoacyl-tRNA hydrolytic domain of phenylalanyl-tRNA synthetase are functionally and evolutionarily divergent. Collectively, these results substantially expand our understanding of the CtdA family, providing new insights into its structure, function, and evolution. Moreover, this work highlights the diverse mechanisms unique to each organism to ensure faithful translation of the genetic code.

biochemistry↗

Two cowpea Rubisco activase isoforms for crop thermotolerance

O_LIA vital crop for sub-Saharan Africa, cowpea productivity is threatened by climate change, including increased heatwave intensity, duration, and frequency. Rubisco activase (Rca) is a key molecular chaperone that maintains Rubisco activity and due to its thermal sensitivity is a key target for improving crop climate resilience. C_LIO_LIWe identified and characterised four Rca isoforms in Vigna unguiculata (L.) Walp. cv. IT97K-499-35 in vitro: Rca1{beta}, Rca8, Rca10 and Rca10{beta}. Cowpea leaf and plant growth traits were also investigated during a 5-day +10 {degrees}C heatwave that included assessment of Rubisco and Rca activity in leaf extracts, and relative changes in the abundance of the four Rca isoforms. C_LIO_LICowpea Rca10 and Rca10{beta} had higher thermal maxima, broader thermal optima and higher rates of ATP hydrolysis and Rubisco reactivation in vitro. In the absence of water deficit, the heatwave caused only mild effects, including increases in leaf temperature and expression of Rca10, small decreases in Rubisco activity and activation state, and unaltered temperature response of Rubisco activation by Rca. C_LIO_LIThe superior Rca10 and Rca10{beta} isoforms offer the prospect of enhancing the thermotolerance of cowpea and other crops in anticipation of more extreme future heatwaves. C_LI

plant biology↗

GLUT5 armouring enhances adoptive T cell therapy anti-tumour activity under glucose-limiting conditions

Cancer immunotherapy with engineered T cells has become a standard treatment for certain hematologic cancers. However, clinical trial outcomes for solid tumours are significantly lagging. A primary challenge in solid tumours is the lack of essential metabolites in the tumour microenvironment, such as glucose, due to poor vascularization and competition with tumour cells. To address this, we modified T cells to use fructose as an alternative energy source by introducing ectopic GLUT5 expression. We show that "GLUT5-armored" T cells, engineered with either chimeric antigen receptors (CARs) or an ectopic T cell receptor (TCR), achieve enhanced anti-tumour activity in low-glucose environments in both in vitro and in vivo models. This straightforward modification is compatible with current clinical approaches and may improve the efficacy of T cell therapies for solid tumours. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=117 SRC="FIGDIR/small/633365v1_ufig1.gif" ALT="Figure 1"> View larger version (62K): org.highwire.dtl.DTLVardef@15e10e0org.highwire.dtl.DTLVardef@a5f029org.highwire.dtl.DTLVardef@3a32daorg.highwire.dtl.DTLVardef@e09ff9_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Cryo-EM structures of PP2A:B55-Eya3 and PP2A:B55-p107 define PP2A:B55 substrate recruitment

The phosphoprotein phosphatase (PPP) family of ser/thr phosphatases are responsible for the majority of all ser/thr dephosphorylation in cells. However, unlike their kinase counterpart, they do not achieve specificity via phosphosite recognition sequences, but instead bind substrates and regulators using PPP-specific short linear and/or helical motifs (SLiMs, SHelMs). Protein phosphatase 2A (PP2A) is a highly conserved PPP that regulates cell signaling and is a tumor suppressor. Here, we investigate the mechanisms of substrate and regulator recruitment to the PP2A:B55 holoenzyme to define how substrates and regulators engage B55 and understand, in turn, how these interactions direct phosphosite dephosphorylation. Our cryo-EM structures of PP2A:B55 bound to p107 (substrate) and Eya3 (regulator), coupled with biochemical, biophysical and cell biology assays, show that while B55 associates using a common set of interaction pockets, the mechanisms of substrate and regulator binding can differ substantially. This shows that B55-mediated substrate recruitment is distinct from that observed for PP2A:B56 and other PPPs. It also allowed us to identify the core B55 recruitment motif in Eya3 proteins, a sequence we show is conserved amongst the Eya family. Finally, using NMR-based dephosphorylation assays, we also showed how B55 recruitment directs PP2A:B55 fidelity, via the selective dephosphorylation of specific phosphosites. Because of the key regulatory functions of PP2A:B55 in mitosis and DNA damage repair, these data provide a roadmap for pursuing new avenues to therapeutically target this complex by individually blocking a subset of regulators that use different B55 interaction sites.

biochemistry↗

An RGB based deep neural network for high fidelity Fusarium head blight phenotyping in wheat

Fusarium head blight (FHB) in wheat is an economically important disease, which can cause yield losses exceeding 50% and the causal pathogen that infects spikes produces harmful mycotoxins. Breeding for host resistance remains the most effective disease control method; but time, labor, and human subjectivity during disease scoring limits selection advancements. In this study we describe an innovative, high-throughput phenotyping rover for capturing in-field RGB images and a deep neural network pipeline for wheat spike detection and FHB disease quantification. The image analysis pipeline successfully detects wheat spikes from images under variable field conditions, segments spikes and diseased tissue in the spikes, and quantifies disease severity as the region of intersection between spike and disease masks. Model inferences on an individual spike and plot basis were compared to human visual disease scoring in the field and on imagery for model evaluation. The precision and throughput of the model surpassed traditional field rating methods. The accuracy of FHB severity assessments of the model was equivalent to human disease annotations of images, however individual spike disease assessment was influenced by field location. The model was able to quantify FHB in images taken with different camera orientations in an unseen year, which demonstrates strong generalizability. This innovative pipeline represents a breakthrough in FHB phenotyping, offering precise and efficient assessment of FHB on both individual spikes and plot aggregates. The model is robust to different conditions and the potential to standardize disease evaluation methods across the community make it a valuable tool for studying and managing this economically significant fungal disease.

plant biology↗

Cryo-EM structures of PP2A:B55-FAM122A and PP2A:B55-ARPP19

Progression through the cell cycle is controlled by regulated and abrupt changes in phosphorylation.1 Mitotic entry is initiated by increased phosphorylation of mitotic proteins, a process driven by kinases,2 while mitotic exit is achieved by counteracting dephosphorylation, a process driven by phosphatases, especially PP2A:B55.3 While the role of kinases in mitotic entry is well-established, recent data have shown that mitosis is only successfully initiated when the counterbalancing phosphatases are also inhibited.4 For PP2A:B55, inhibition is achieved by the two intrinsically disordered proteins (IDPs), ARPP19 (phosphorylation-dependent)6,7 and FAM122A5 (inhibition is phosphorylation-independent). Despite their critical roles in mitosis, the mechanisms by which they achieve PP2A:B55 inhibition is unknown. Here, we report the cryo-electron microscopy structures of PP2A:B55 bound to phosphorylated ARPP19 and FAM122A. Consistent with our complementary NMR spectroscopy studies both IDPs bind PP2A:B55, but do so in highly distinct manners, unexpectedly leveraging multiple distinct binding sites on B55. Our extensive structural, biophysical and biochemical data explain how substrates and inhibitors are recruited to PP2A:B55 and provides a molecular roadmap for the development of therapeutic interventions for PP2A:B55 related diseases.

biochemistry↗

PP2A/B55α substrate recruitment as defined by the retinoblastoma-related protein p107

Protein phosphorylation is a reversible post-translation modification essential in cell signaling. This study addresses a long-standing question as to how the most abundant serine/threonine Protein Phosphatase 2 (PP2A) holoenzyme, PP2A/B55, specifically recognizes substrates and presents them to the enzyme active site. Here, we show how the PP2A regulatory subunit B55 recruits p107, a pRB-related tumor suppressor and B55 substrate. Using molecular and cellular approaches, we identified a conserved region 1 (R1, residues 615-626) encompassing the strongest p107 binding site. This enabled us to identify an "HxRVxxV619-625" short linear motif (SLiM) in p107 as necessary for B55 binding and dephosphorylation of the proximal pSer-615 in vitro and in cells. Numerous B55/PP2A substrates, including TAU, contain a related SLiM C-terminal from a proximal phosphosite, "p[ST]-P-x(5-10)-[RK]-V-x-x-[VI]-R". Mutation of conserved SLiM residues in TAU dramatically inhibits dephosphorylation by PP2A/B55, validating its generality. A data-guided computational model details the interaction of residues from the conserved p107 SLiM, the B55 groove, and phosphosite presentation. Altogether these data provide key insights into PP2A/B55 mechanisms of substrate recruitment and active site engagement, and also facilitate identification and validation of new substrates, a key step towards understanding PP2A/B55[<] role in multiple cellular processes.

biochemistry↗

Open Natural Products Research: Curation and Dissemination of Biological Occurrences of Chemical Structures through Wikidata

Contemporary bioinformatic and chemoinformatic capabilities hold promise to reshape knowledge management, analysis and interpretation of data in natural products research. Currently, reliance on a disparate set of non-standardized, insular, and specialized databases presents a series of challenges for data access, both within the discipline and for integration and interoperability between related fields. The fundamental elements of exchange are referenced structure-organism pairs that establish relationships between distinct molecular structures and the living organisms from which they were identified. Consolidating and sharing such information via an open platform has strong transformative potential for natural products research and beyond. This is the ultimate goal of the newly established LOTUS initiative, which has now completed the first steps toward the harmonization, curation, validation and open dissemination of 750,000+ referenced structure-organism pairs. LOTUS data is hosted on Wikidata and regularly mirrored on https://lotus.naturalproducts.net. Data sharing within the Wikidata framework broadens data access and interoperability, opening new possibilities for community curation and evolving publication models. Furthermore, embedding LOTUS data into the vast Wikidata knowledge graph will facilitate new biological and chemical insights. The LOTUS initiative represents an important advancement in the design and deployment of a comprehensive and collaborative natural products knowledge base. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=61 SRC="FIGDIR/small/433265v3_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@184136borg.highwire.dtl.DTLVardef@16e0d6org.highwire.dtl.DTLVardef@30ae1org.highwire.dtl.DTLVardef@1bf4825_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

The interaction of p38 with its upstream kinase MKK6

Mitogen-activated protein kinase (MAPK; p38, ERK and JNK) cascades are evolutionarily conserved signaling pathways that regulate the cellular response to a variety of extracellular stimuli, such as growth factors and interleukins. The MAPK p38 is activated by its specific upstream MAPK kinases, MKK6 and MKK3. However, a comprehensive molecular understanding of how these cognate upstream kinases bind and activate p38 is still missing. Here, we combine NMR spectroscopy and isothermal titration calorimetry to define the binding interface between full-length MKK6 and p38. We show that p38 engages MKK6 not only via its hydrophobic docking groove, but also helix F, a secondary structural element that plays a key role in organizing the kinase core. We also show that, unlike MAPK phosphatases, the p38 conserved docking (CD) site is much less affected by MKK6 binding. Finally, we demonstrate that these interactions with p38 are conserved independent of the MKK6 activation state. Together, our results reveal differences between specificity markers of p38 regulation by upstream kinases, which do not effectively engage the CD site, and downstream phosphatases, which require the CD site for productive binding.

biophysics↗

1H, 15N and 13C sequence specific backbone assignment of the MAP Kinase Binding Domain of the Dual Specificity Phosphatase 1 and its interaction with the MAPK p38

The sequence-specific backbone assignment of the mitogen-activated protein kinase (MAPK) binding domain of the dual-specificity phosphatase 1 (DUSP1) has been accomplished using a uniformly [13C,15N]-labeled protein. These assignments will facilitate further studies of DUSP1 in the presence of inhibitors/ligands to target MAPK associated diseases and provide further insights into the function of dual-specificity phosphatase 1 in MAPK regulation.

biophysics↗

Inbreeding depression causes reduced fecundity in Golden Retrievers

Inbreeding depression has been demonstrated to impact vital rates, productivity, and performance in many domestic species. Many in the field have demonstrated the value of genomic measures of inbreeding compared to pedigree-based estimates of inbreeding; further, standardized, high-quality phenotype data on all individuals is invaluable for longitudinal analyses of a study cohort. We compared measures of reproductive fitness in a small cohort of Golden Retrievers enrolled in the Golden Retriever Lifetime Study (GRLS) to a genomic measurement of inbreeding, FROH. We demonstrate a statistically significant negative correlation between fecundity and FROH.This work sets the stage for larger scale analyses to investigate genomic regions associated with fecundity and other measures of fitness.

genomics↗

Development of new food-sharing relationships among nonkin vampire bats

In an individualized animal society, social bonds can foster cooperation and enhance survival and reproduction. Cooperative bonds often exist among kin, but nonkin can also develop high-investment cooperative bonds that share similarities with human friendship. How do such bonds form? One theory suggests that strangers should test the waters of a new relationship by making small initial cooperative investments and gradually escalating them with good partners. This raising-the-stakes strategy is demonstrated by human strangers in short-term economic games, but it remains unclear whether it applies to helping in a natural long-term social bond. Here we show evidence that unfamiliar vampire bats (Desmodus rotundus) selectively escalate low-cost investments in allogrooming before developing higher-cost food-sharing relationships. We introduced females from geographically distant sites in pairs or groups and observed that bats established new reciprocal grooming relationships, and that increasing grooming rates predicted the occurrence of first food donations, at which point grooming rates no longer increased. New food-sharing relationships emerged reciprocally in 14% of female pairs, typically over 10-15 months, and developed faster when strangers lacked alternative familiar partners. A gradual grooming-to-sharing transition among past strangers suggests that raising the stakes might be more evident when tracking multiple cooperative behaviours as new relationships form, rather than measuring a single behavior in an established relationship. Raising the stakes could play a similar underappreciated role across a broader spectrum of social decisions with long-term consequences, such as joining a new social group or forming a long-term pair-bond. Significance statementVampire bats form long-term cooperative social bonds that involve reciprocal food sharing. How do two unrelated bats go from being strangers to having a high-investment food-sharing relationship? We introduced unfamiliar bats and found evidence that low-cost grooming paves the way for higher-cost food donations. Food sharing emerged in a reciprocal fashion and it emerged faster when two strangers did not have access to their original groupmates. The bats that formed new food-sharing relationships had a history of escalating reciprocal grooming up until the food sharing began. Our finding that unfamiliar nonkin vampire bats appear to gradually and selectively transition from low-cost to high-cost cooperative behaviors is the first evidence that nonhuman individuals raise the stakes when forming new cooperative relationships.

animal behavior and cognition↗