Biology subjects

Wang, J.

Publications and source records attributed to Wang, J..

At least 343 records · Page 19Linked to original sources

bioRxiv2026

The world's first cloned golden wild yak via interspecific SCNT: 4800m donor origin and 4200m vitrified blastocyst transfer

Primarily inhabiting the harsh, high-altitude environment of the Qiangtang National Nature Reserve exceeding 5,000 meters above the sea (m.a.s.l.), the golden wild yak is critically endangered, with fewer than 300 individuals remaining in the world, a situation exacerbated by the significant challenges of conducting research and conservation of their genetic resources. Somatic cell nuclear transfer (SCNT) can be an effective method for their preservation, but facing several obstacles in this context, including the hypoxic stress at high altitude that impairs embryonic development due to in vitro manipulation, and constraints of long-distance embryo transport. In the present study, the ear tissue was collected from a childhood male golden wild yak at Xizang Geye Wildlife Rescue Station (4800 m.a.s.l.) and send to Institute of Animal Science at Beijing to derive fibroblast cells. Using fibroblast cells of the golden wild yak as nuclear donors, and bovine oocytes from a local slaughterhouse at Beijing as recipients, the interspecific SCNT (iSCNT) embryos were generated and in vitro developed to blastocysts. To maintain the embryonic viability after long-distance transportation from Beijing to Xizang, iSCNT blastocysts were subjected to cryopreservation by vitrification method. Thawing of vitrified iSCNT blastocysts were completed at Xizang Dangxiong Yak Breeding Innovation Base (4200 m.a.s.l.), and transferred into the uterine horn of domestic yaks. 257 days after blastocyst transfer, a cloned golden wild yak was successfully harvested on January 10, 2026. This work demonstrates, for the first time, that interspecies somatic cell nuclear transfer can successfully produce a cloned offspring under extreme conditions, spanning 4800 m.a.s.l. donor origin, long-distance vitrified embryo transportation, and high-altitude blastocyst transfer at 4200 m.a.s.l., establishing a viable strategy for conserving critically endangered high-altitude species.

genetics↗

bioRxiv2026

ALPINE: A Scalable Pipeline for Comprehensive Classification of Gene-Editing Outcomes from Long-Read Amplicon Sequencing

CRISPR genome editing has enabled precise genetic modification for gene and cell therapies, but edits often produce heterogeneous on-target outcomes, including homology-directed repair (HDR) knock-ins, DNA repair template integrations, and structural variants. Existing tools are frequently limited to short reads or lack viral vector-specific integration categories needed for therapeutic development. Here, we present ALPINE (Amplicon Long-read Pipeline for INtegration Evaluation), a scalable and reproducible pipeline for classifying and quantifying gene-editing outcomes from long-read amplicon sequencing. ALPINE classifies reads into 10+ categories, including DNA repair vector integration subtypes, and performs variant calling near the gene-edited site with batch, multi-sample reporting. Uniquely, ALPINE can distinguish between cells treated with multiple DNA repair vectors and identify distinct molecular features, such as inverted terminal repeats (ITRs), enabling comprehensive characterization of complex gene editing outcomes. Benchmarking on simulated datasets showed high accuracy, and application to edited T cell samples demonstrated comprehensive gene-editing outcome profiling. Supplementary data are available online. AvailabilityALPINE is implemented in Python and distributed as Docker containers with Common Workflow Language (CWL) support for cloud deployment. The pipeline is available under MIT license at https://github.com/Maggi-Chen/ALPINE.

bioinformatics↗

bioRxiv2026

A Conserved Metabolic Oxidative Axis Underlies Immune Cell Cryo-vulnerability

Immunotherapy has emerged as a transformative approach for treating cancer and other diseases, yet its widespread deployment requires effective cryopreservation strategies to enable scalable global distribution. However, many immune cell types remain acutely vulnerable to freeze-thaw stress, and the underlying mechanisms of this cryo-vulnerability are not well defined. In this study, we combined metabolic flux analysis, ROS quantification, lipidomics, and preclinical xenograft models to investigate how metabolic state influences cryopreservation outcomes. We found that immune cell activation induces a metabolic shift characterized by elevated glucose utilization and excessive ROS production, leading to profound post-thaw loss of viability and function, as demonstrated by a [~]25% survival rate in natural killer (NK) cells. Targeted pretreatments--including inhibitors of glucose metabolism, antioxidants, and suppression of lipid peroxidation--restored post-thaw recovery to nearly 90% while preserving effector activity and antitumor efficacy. Similar protective effects were observed across {beta} T cells, {gamma}{delta} T cells, and macrophages, defining a conserved metabolic-oxidative pathway of cryo-vulnerability and offering applicable strategies to enhance immune cell preservation.

immunology↗

bioRxiv2026

Cryo-EM structures of photosystem I with alternative quinones reveals new insight into cofactor selectivity

Quinones are an integral component of electron transfer processes in photosynthetic and mitochondrial respiratory proteins. One such photosynthetic protein, Photosystem I, is an essential photooxidoreductase found in all oxygenic phototrophs. To better understand quinone chemistry and to form a basis for protein engineering, the menB gene in the model cyanobacterium Synechocystis sp. PCC 6803 was interrupted, blocking the biosynthesis of phylloquinone and causing it to be replaced by exchangeable plastoquinone-9 in the A1A and A1B quinone-binding sites of Photosystem I. This genetic variant has been instrumental in bioenergy research, enabling incorporation of a range of substituted and isotopically labeled quinones. Despite numerous valuable studies, the interpretation of biophysical data has been limited by a lack of structural data. To address this, we present the high-resolution cryo-EM structures of Photosystem I from the {Delta}menB variant containing (a) exchangeable plastoquinone-9 and (b) exogenously added 2-ethyl-1,4-napthoquinone at 1.90- and 2.05-[A] resolution, respectively. Unexpectedly, the quinones in the A1A and A1B sites of Photosystem I, previously believed to have similar binding affinities, are found to be asymmetric in their ability to bind and exchange plastoquinone-9. This work reveals new and important insight into the molecular basis for Photosystem I activity in the {Delta}menB variant, the power of metabolic plasticity to maintain protein stability, and the requirement for protein instability to facilitate ligand exchange.

biochemistry↗

bioRxiv2026

Complete genome-derived metabolic interactions reveal impact of gut ecology on human health

Metabolic interactions govern gut microbiome assembly, yet functional rules remain obscured by genomic incompleteness and fragmentation. Here, we leverage 1,150 complete genomes to construct genome-scale metabolic models, demonstrating that draft genomes introduce systemic artifacts missing critical transport functions. We observe that metabolic interactions are intrinsically driven by genomic traits and niche specialization rather than random association. We stratifies strains into four ecological groups, including active players, resource predators, utilizers, and contributors, each of which exhibits distinct metabolite exchange/competition signatures. This functional architecture is strongly coupled to secondary metabolism. Applying this framework to inflammatory bowel disease revealed distinct temporal dynamics of ecological groups across disease subtypes, where group-specific dysbiosis predicted clinical phenotypes better than whole community profiles. Furthermore, integrating metabolic and co-occurrence networks to identify keystone for classifiers as features significantly improved disease diagnosis. Our study provides novel insights into gut metabolic interaction landscape, bridging the gap between genomic potential and clinical utility for precision microbiome therapeutics.

microbiology↗

bioRxiv2026

A genome-wide CRISPR Screening identifies targets that drive Tolerogenic Dendritic Cells

Tolerogenic dendritic cells (TolDCs) are essential for immune tolerance and offer promise for treating autoimmune diseases. Despite the clinical evidence of their therapeutic potential, the key molecular pathways guiding their differentiation and tolerogenic phenotype remain elusive due to complex interactions identified in functional assays. Here we investigated the molecular profiles and regulatory programs underlying the functional status of tolerogenic dendritic cell populations in response to known tolerizing agents. We identified CD86 as a consistent and robust marker downregulated in tolerogenic state. Using CD86 blocking antibodies or CRISPR-mediated gene inactivation we demonstrated that CD86 is functionally required for TolDC-mediated suppression of T cell proliferation and cytokine secretion, establishing CD86 as both a consensus phenotypic and functional screening marker and a mechanistic regulator of tolerance. Leveraging CD86 as a scalable readout, we performed a pooled genome-wide CRISPR-Cas9 knockout screen to identify regulators of TolDC function. This approach uncovered UBE2L6 as a novel modulator that promotes the tolerogenic phenotype and restricts TolDC-mediated T cell activation. Mechanistically, UBE2L6 deficiency leads to coordinated upregulation of ISG15 and USP18, indicating a possible ISGylation-dependent pathway regulating CD86 expression and tolerogenic function. Together, this study identifies pathways that can be targeted to promote immune tolerance in immune-mediated inflammatory diseases. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=105 SRC="FIGDIR/small/713621v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@4c3636org.highwire.dtl.DTLVardef@17b27e5org.highwire.dtl.DTLVardef@783d95org.highwire.dtl.DTLVardef@1337ce_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2026

Complete Genomes of Cultivated Gut Bacteria Reveal Mobile Genetic Element-Driven Functional Diversity with Therapeutic Implications

Cultivated bacterial isolates are essential for elucidating gut microbiota functions, yet reliance on fragmented draft genomes and limited mobile genetic element (MGE) annotation has constrained high-resolution analyses. Here, we present the Cultivated Complete Genome Reference (CCGR), a comprehensive compendium of 1,150 fully circularized human gut bacterial genomes. Overcoming draft limitations, CCGR achieves chromosome-scale resolution and reveals distinct evolutionary strategies shaping gene topological organization in response to bacterial growth. Furthermore, we decode the dynamic landscape of MGEs, demonstrating that broad-host-range phages follow strict positional patterns by integrating at conserved chromosomal hotspots flanked by nutrient transporter loci. Importantly, we provide mechanistic in vivo evidence that accessory genomic elements drive strain-level functional heterogeneity. Specifically, the plasmid-encoded scrK gene in Levilactobacillus brevis involved in fructose catabolism is required to mitigate fructose-diet-exacerbated colitis. Collectively, CCGR establishes a framework linking chromosomal architecture and MGEs to bacterial evolution and host health, enabling precise, strain-resolved functional investigations.

microbiology↗

bioRxiv2026

Unraveling Viral peptide-G4 Interactions: the NS3 Protease Domain of Yellow Fever Virus Binds G-Quadruplexes with High Specificity and Affinity

G-quadruplexes (G4s) are critical nucleic acid secondary structures that play pivotal roles in regulating gene expression. In this study, we conducted a proteome-wide in silico analysis across multiple viruses causing hemorrhagic fevers to identify candidate proteins containing a conserved G4-binding motif. Four peptides belonging to Marburg, Ebola, Hantaan and Yellow fever viruses were shown to bind to G4 in vitro. We selected the NS3 protease domain of Yellow Fever virus for further validation. Biochemical assays demonstrated that the NS3 protease domain binds G4 structures with high specificity and affinity, particularly favoring the parallel conformation. Molecular docking and simulations further revealed that the NS3 protease domain interacts with the terminal G-tetrads and loop regions of G4 via key residues, including PHE40, adopting an insertion and stacking composite binding mode. These findings expand our understanding of virus - G4 interactions and offer novel potential targets for G4-based antiviral strategies. Bullet points- We screened viruses causing hemorrhagic fevers for potential G4-binding peptides. - Four peptides belonging to Marburg, Ebola, Hantaan and Yellow fever viruses were shown to bind to G4 in vitro. - Biochemical assays demonstrated that the NS3 protease domain of YFV binds G4 structures with high specificity and affinity.

biophysics↗

bioRxiv2026

Gq-pathway activation in hippocampal CA1 astrocytes rescues ischemia-induced memory deficits and synaptic plasticity

More than 30% of stroke survivors develop post-stroke cognitive impairment (PSCI), for which current neuron-centric therapies not only lack cellular specificity but also carry risks of adverse effects such as epilepsy. Here, we explore the therapeutic potential of astrocytes by chemogenetically activating Gq signaling in hippocampal astrocytes, which rescues memory deficits and synaptic plasticity impairments in a mouse model of ischemic stroke. Gq activation restores dendritic complexity, spine density, and long-term potentiation in hippocampal CA1 neurons. Fiber photometry further reveals that astrocytic Ca{superscript 2} signals precede neuronal activity by 600 ms during novel environment exploration, indicating that astrocytes prime memory encoding. In contrast, Gi pathway activation induces pathological neuronal hyperactivity without cognitive improvement. These findings establish that astrocytes regulate post-stroke recovery through pathway-specific calcium signaling and uncover a previously unknown temporal hierarchy in astrocyte-neuron communication during memory processing, offering a new glia-targeted strategy to overcome the limitations of current neuromodulation approaches for PSCI.

neuroscience↗

bioRxiv2026

Two ancient nuclear lineages and divergent reproductive strategies drive global success of the wheat stripe rust pathogen

Understanding the mechanisms for generating and maintaining plant pathogen genetic diversity is essential for predicting disease emergence and safeguarding the food supply. Here, using population-genomic analyses of 507 global isolates of the wheat stripe rust fungus Puccinia striiformis f. sp. tritici, we uncover two nuclear lineages that diverged before modern agriculture and reinforced during wheat domestication. Distinct lineage combinations underpin three major global populations: two predominantly sympatric, homozygous lineages (nuclA-nuclA and nuclB-nuclB) and a globally dominant heterokaryotic lineage (nuclA-nuclB) of hybrid origin. This hybrid population evolves largely clonally, accompanied by recurrent somatic nuclear exchange. Identical virulence traits can arise independently via somatic nuclear exchange in clonal populations or through sexual recombination in recombining populations, revealing convergent evolutionary routes to adaptation. Together, our results provide a transformative nuclear-level perspective linking deep evolutionary history and reproductive strategy to the global success of cereal rust fungi.

genetics↗

bioRxiv2026

ChironRNA: Steric Clashes Resolution in RNA Structures via E(3)-Equivariant Diffusion

Due to the limited resolution of experimental data, many determined RNA structures contain physically implausible geometries, such as severe steric clashes and missing atoms. Resolving these defects during RNA structure refinement remains a fundamental challenge. Structure dictates the function, so the geometric accuracy of RNA structure is critical for understanding biological mechanisms. However, traditional algorithms for correction have limitations because of the complexity of RNA structures. We propose ChironRNA, an all-atom diffusion model with E(3)-equivariant graph neural networks to perform RNA refinement by resolving steric clashes and completing missing atoms. In ChironRNA, we adopt a hierarchical approach, including both an all-atom diffusion model and a coarse-grained diffusion model where each nucleotide is represented by a five-point representation. Our pipeline consists of two stages: a training stage and a generation stage. The diffusion model regenerates clashing nucleotide atoms step by step by removing the noise predicted by EGNN. ChironRNA achieves an 80% clash reduction on more than 80% of the test set. It performs better on structures of less than 200 nucleotides, resulting in a high percentage of cases having over 80% clash reduction rate and 100% atom reconstruction rate. Our results demonstrate that ChironRNA successfully resolves steric clashes and rebuilds missing atoms with high precision, offering a robust solution where traditional fine-tuning or enumerative approaches fail.

biophysics↗

bioRxiv2026

PalmaClust: A graph-fusion framework leveraging the Palma ratio for robust ultra-rare cell type detection in scRNA-seq data

MotivationSingle-cell RNA sequencing (scRNA-seq) is routinely used to build atlases of tissues, resolve developmental trajectories, and characterize disease microenvironments. Yet many biologically and clinically meaningful populations--including transient progenitors, therapy-resistant tumor subclones, and antigen-specific lymphocytes--occur at very low frequencies (<1%) and are easily missed by standard clustering pipelines. Existing approaches often require extensive manual curation, rely on known marker genes, or trade sensitivity for unacceptable false positive rates due to the insensitivity of metrics like the Gini index to heavy-tailed distributions. A scalable, statistically grounded method is needed to sensitively detect rare populations while providing calibrated confidence and interpretable molecular signatures. ResultsWe present PalmaClust, a graph-fusion clustering framework that repurposes Palma ratio--a tail-sensitive inequality metric in sociology--to identify marker genes driven by extreme sparsity. PalmaClust constructs and fuses multiple K-Nearest Neighbor (KNN) graphs derived from complementary gene-selection statistics including the Palma ratio, Gini index, and Fano factor. It employs a local refinement strategy that re-prioritizes Palma-ranked genes within parent clusters. Benchmarking across diverse public scRNA-seq datasets confirms that PalmaClust consistently outperforms state-of-the-art baselines, improving rare-class F1 scores by at least 20% (absolute) while maintaining high global clustering stability. Further studies demonstrate that the Palma ratio-derived graph layer is essential for capturing ultra-rare signatures that other views miss. Availabilityhttps://github.com/wan-mlab/PalmaClust.

bioinformatics↗

bioRxiv2026

PPa1 insufficiency drives lysosomal storage disease and inflammatory macrophage expansion in the bone marrow.

Inorganic pyrophosphatase-1 (PPa1) is an essential enzyme proposed to limit accumulation of the ubiquitous metabolic byproduct inorganic pyrophosphate, yet its role beyond a general housekeeping function remains poorly understood. We generated viable hypomorphic alleles of PPa1 through random mutagenesis in mice and unexpectedly found that PPa1 insufficiency causes a lysosomal storage disease of the bone marrow. Mutant mice developed impaired hematopoiesis and defective skeletal mineralization in a hematopoietic-intrinsic manner. The bone marrow was infiltrated by glycolipid-laden macrophages that were marked by high Spp1 and CD14 expression, blunted lysosomal acidification, metabolic stress, and a distinct inflammatory transcriptional program. Consistent with a lysosomal storage defect, PPa1-deficient bone marrow accumulated elevated levels of long-chain glucosyl-sphingolipids and sphingosine. These findings identify PPa1 as a previously unrecognized regulator of lysosomal function in myeloid cells that restrains inflammatory macrophage expansion and prevents bone marrow lysosomal storage pathology.

immunology↗

bioRxiv2026

A Long-Context Generative Foundation Model Deciphers RNA Design Principles

Programmable design of RNA sequences with defined functions remains a central challenge in biology. Despite recent advances, existing RNA generative models lack robust controllable design capabilities and are constrained by short context windows, limiting their capacity to model the complex evolutionary manifold of full-length transcripts. Here, we introduce EVA (Evolutionary Versatile Architect), a long-context generative RNA foundation model trained on over 114 million full-length RNA sequences spanning the full breadth of evolutionary diversity. By integrating a Mixture-of-Experts architecture with an 8,192-token context window, EVA learns a unified, evolution-consistent representation of RNA sequence space that captures both global transcript structure and fine-grained functional elements. EVA establishes a unified framework for a broad spectrum of RNA design tasks within a single architecture, ranging from mutational fitness prediction to context-aware sequence engineering. We demonstrate EVAs versatility through the de novo design and targeted optimization of diverse RNA classes, including tRNAs, aptamers, and CRISPR guide RNAs, as well as mRNA and circular RNA therapeutics. In systematic evaluations, EVA achieves state-of-the-art performance across seven of nine established benchmarks, improving structural modeling accuracy by up to an order of magnitude compared to existing approaches. EVA is fully open-source at the following repository: https://github.com/GENTEL-lab/EVA.

molecular biology↗

bioRxiv2026

Encounter-state over-anchoring governs productive PETase binding on PET surfaces

Polyethylene terephthalate (PET) hydrolysis by Ideonella sakaiensis PETase (IsPETase) begins at a heterogeneous solid-liquid interface, yet the molecular basis of productive surface recognition remains poorly resolved. Here, we combined a Martini 3 coarse-grained PET model with G[o]Martini protein dynamics to investigate IsPETase binding to an extended PET surface. A four-state kinetic model, comprising unbound, encounter, docked, and pre-catalytic states, shows that productive binding is not limited by adsorption itself, but by a post-adsorption re-registration step that converts surface-bound encounter complexes into productively aligned configurations. The simulations reveal a stage-dependent role of conformational flexibility: flexible surface loops facilitate early capture, whereas excessive flexibility promotes misregistered hydrophobic contacts, over-stabilizes non-productive encounter states, and lowers the overall probability of productive commitment. Analysis of productive trajectories further identifies three microscopic reorientation modes by which the enzyme reaches the pre-catalytic state after adsorption. Comparative simulations of engineered PETase variants uncover a flexibility-driven speed-yield trade-off, in which increased flexibility accelerates successful binding events but reduces productive yield through encounter-state over-anchoring. Guided by this mechanism, we formulated a landscape-based design strategy that either weakens encounter-specific anchors or reinforces product-like contacts, leading to mutations that improve productive-binding yield. These results identify post-adsorption alignment as the key kinetic bottleneck in PETase surface recognition and provide a mechanistic framework for designing enzymes that operate at heterogeneous polymer interfaces.

biophysics↗

bioRxiv2026

Indazolone-Based Molecular Glue Degraders as a Tunable Platform for Reprogramming Cereblon Substrate Specificity

Molecular glue degraders (MGDs) represent a transformative modality in drug discovery, with the cereblon (CRBN)-MGD axis offering a strategic gateway to systematically address the historically undruggable proteome. However, the limited chemical diversity of existing CRBN-mediated MGDs has constrained the full exploration of the degradable landscape. Here, we introduce an innovative indazolone-based platform that extends beyond canonical isoindolinone-glutarimide scaffolds, significantly expanding the accessible chemical space for CRBN modulation. Guided by mechanistic insights into CRBN-MGD complex conformational plasticity, we rationally designed indazolone architectures as novel CRBN ligands, developing diverse MGDs with potent ligase binding and exceptional substrate programmability. This platform enables fine-tuned control over neo-substrate recognition, supporting diverse degradation profiles ranging from the broad targeting of critical proteins (IKZF1/3, ZFP91, and LIMD1) to the exquisitely selective degradation of CK1 and IKZF2. Beyond expanding the chemical landscape for intractable targets, our indazolone-based MGDs exhibit favorable pharmacokinetic properties, offering robust promise for therapeutic development. Collectively, the remarkable tunability and exceptional neo-substrate programmability of the indazolone-based platform provide a highly transformative blueprint for next-generation MGD discovery, enabling systematic exploration of the vast CRBN-accessible proteome. Furthermore, as superior CRBN ligands, this platform holds immense potential for PROTAC discovery to systematically address previously intractable targets.

biochemistry↗

bioRxiv2026

The muscle atrophic phenotype of MuSK myasthenia gravis: Insights from a preclinical rat model

Myasthenia gravis with muscle-specific kinase antibodies (MuSK-MG) is an autoimmune disorder marked by neuromuscular junction (NMJ) disruption and selective muscle atrophy, yet its intrinsic myocellular mechanisms remain unclear. Using a rat model generated by active immunization with the N-terminal MuSK60 peptide, we characterized muscle pathology, NMJ morphology, and whole-muscle proteome remodeling across anatomically distinct skeletal muscles. Anti-MuSK rats developed seropositivity, body-mass loss, fragmented and denervated NMJs, and pronounced atrophy restricted to slow-twitch/type I fibers, particularly in the soleus muscle. Quantitative proteomics identified extensive muscle-specific alterations, most prominent in soleus, with fewer in diaphragm and sternohyoideus. Gene set enrichment analysis revealed coordinated downregulation of mitochondrial, ribosomal, and myosin-complex proteins in soleus, partial mitochondrial involvement in diaphragm, and compensatory upregulation of translational and proteasomal pathways in diaphragm. Correlation analysis linked soleus mass loss to elevated abundance of ubiquitin-proteasome and calcium-handling proteins, implicating proteolytic and bioenergetic stress mechanisms. The consistent upregulation of NCAM1 and MUSTN1 suggests generalized myocellular responses to MuSK dysfunction. Together, these data demonstrate that MuSK autoimmunity elicits fiber-type-selective atrophy and profound proteome remodeling beyond NMJ impairment, highlighting disrupted mitochondrial and translational homeostasis as central features of the MuSK-MG muscle phenotype.

molecular biology↗

bioRxiv2026

Mechanistic insight into the phosphorylation of ERK by MEK

The RAS-RAF-MEK-ERK cascade regulates critical cellular processes such as cell proliferation and differentiation, with its dysregulation driving over 30% of human cancers. However, the active state of MEK and molecular mechanisms of ERK phosphorylation remain unclear. Here, we report the cryo-EM structures of phosphorylated MEK1 (pMEK1) bound to unphosphorylated and monophosphorylated ERK1. These structures reveal that pMEK1 interacts with ERK1 via three interfaces: first between MEK1 N-terminal peptide and ERK1 D-site recruitment site (DRS), second between MEK1 C-lobe and ERK1 F-site recruitment site (FRS), and third between MEK1 activation loop and ERK1 activation loop. Notably, we identify an unexpected phosphorylation mechanism: pMEK1 catalyzes ERK1 T202 phosphorylation through phosphate transfer from ERK1 Y204. In addition, pMEK1 exhibits phosphatase activity, catalyzing the dephosphorylation of ERK1 Y204. These findings allow us to delineate the catalytic cycle of MEK-catalyzed ERK phosphorylation and provide insights for targeting this oncogenic pathway.

molecular biology↗