Cancer is a complex disease driven by genetic, metabolic, and environmental alterations, whose investigation is often constrained by the limited tractability of mammalian systems. The budding yeast Saccharomyces cerevisiae has emerged as a powerful eukaryotic model to study conserved cellular processes relevant to tumor biology in a simplified and scalable context and as a versatile platform for translational and biotechnological applications. Through genetic manipulation and heterologous expression, yeast allows systematic analysis of human cancer genes and variants, providing quantitative insights into their functional impact. In parallel, yeast reproduces fundamental features of cancer cell metabolism and stress adaptation, offering a controlled system to investigate cellular responses to environmental constraints. The conservation of major DNA repair and autophagy pathways further supports the use of yeast to study genome stability and survival mechanisms in cancer. Beyond its role in basic research, S. cerevisiae represents a scalable platform for anticancer drug discovery, enabling systematic identification of drug targets, resistance mechanisms, and genotype-specific vulnerabilities through high-throughput and engineered strain-based approaches. Continued development of yeast platforms, together with synthetic biology, functional genomics, and advanced genomic technologies, is expected to accelerate therapeutic innovation and improve our understanding of cancer biology. This review discusses recent advances in yeast-based cancer research, highlighting the contribution of engineered yeast platforms to the investigation of oncogenic signaling, metabolic rewiring, stress adaptation, DNA repair, and autophagy, reinforcing the role of yeast at the interface between cancer research and biotechnology.
Pseudomonas aeruginosa is an opportunistic nosocomial pathogen ranked by the World Health Organization (WHO) as a high priority for research and the development of new antimicrobial therapies. To characterize carbapenem-resistant P. aeruginosa (CRPA) isolated from patients at the Hospital de Urgencia Asistencia Pública (HUAP), Chile in 2022, and to evaluate clonal diversity, resistance mechanisms, virulence factors, and clinical associations. We analyzed 126 clinical CRPA isolates using PFGE, PCR screening of resistance and virulence genes, and whole-genome sequencing of 18 representative strains. Sequence types (STs), resistomes, and virulomes were identified, and their associations with severity, length of hospital stay, cost, and patient outcomes were evaluated. Seventy-four percent of isolates originated from critical care units, predominantly from respiratory and tissue samples. PFGE revealed 25 pulsotypes, with L and Y being the predominant types. Overall, 67% of isolates were XDR and 1% PDR. Carbapenemase genes were absent in 115 isolates, while blaVIM and blaKPC were detected in 8 and 3 isolates, respectively. The exoS +/exoU + genotype was identified in 6 isolates; whole-genome sequencing revealed eight distinct STs, including previously described high-risk clones ST654, ST395, and ST274. A resistome analysis revealed diverse aminoglycoside and β -lactam resistance determinants, while a virulome analysis confirmed the presence of exoS +/exoU + in two sequenced isolates. Carbapenem resistance was significantly associated with prolonged hospitalization (median 84 vs. 39 days), greater clinical severity, and substantially higher healthcare resource utilization, reflected by increased DRG (Diagnosis-Related Group) weights, compared with carbapenem-susceptible controls. Our findings highlight the circulation of high-risk P. aeruginosa clones in Chile and underscore the importance of molecular epidemiology in guiding infection control, optimizing antimicrobial therapy, and mitigating the clinical and economic burden of CRPA.
Our immune system is constantly exposed to fungi, but it mounts a consistent, disease-related, and species-specific inflammatory response only against a few fungal species. Most of the current understanding of fungus-host interactions is based on a limited number of strains, hence neglecting the fungal intra-specific genetic and phenotypic diversity. To expand our knowledge of the spectrum of immune responses to pathogenic and non-pathogenic fungi, we compared the cytokine and transcriptional profiles of human monocyte-derived dendritic cells exposed to Aspergillus fumigatus, Candida albicans, Candida parapsilosis, and Saccharomyces cerevisiae strains. The tested species triggered common and species-specific responses, mostly resulting from the different timing of signaling pathways. Faster phagolysosome acidification was observed for pathogenic species. These results highlight the urgency to redraw the boundaries between pathogenicity and commensalism in fungi, shining a spotlight on the timing of the response, rather than solely on the genes triggered by the stimuli.
Sorbic acid is a lipophilic weak acid with fungistatic activity, and it has been widely used as a food preservative, along with its potassium and calcium salts. Although the fungistatic effect of sorbic acid is thought to be primarily due to acidification within fungal cells, the detailed fungistatic mechanism remains unclear. We investigated the effects of sorbic acid on yeast translation in Saccharomyces cerevisiae. At sublethal concentrations (2-4 mM), sorbic acid quickly repressed translation. Conversely, removal of sorbic acid restored translation activity, indicating that the sorbic acid-induced translational repression is reversible. Pronounced translational repression induced by various stress conditions or nutrient starvation is often accompanied by eIF2 α phosphorylation, eIF2B-body and stress granule (SG) formation, and the sequestration of Ded1 (which plays a role in translation initiation as a DEAD-box RNA helicase) into SGs. We found that sorbic acid stress also induces eIF2 α phosphorylation and the sequestration of Ded1 into SGs. In contrast, sorbic acid stress induced the formation of not eIF2B bodies but eIF2B granules, which colocalized with SGs. These results suggest that the functional arrest of translation-related factors, including eIF2 α , eIF2B, and Ded1, correlates strongly with the translational repression in the presence of sorbic acid. Notably, Gcn2 deficiency delayed translational repression and SG formation, and significantly suppressed eIF2B granule formation, suggesting the involvement of Gcn2 in these stress responses during sorbic acid stress. Our findings provide new insights into the physiological effects of sorbic acid on yeast cells, specifically regarding the regulation of translation-related factors.
The Yippee-like (YPEL) proteins are a evolutionarily conserved eukaryotic family implicated in proliferation, senescence, and stress adaptation, yet their molecular functions remain poorly defined. Humans possess five paralogs (YPEL1-YPEL5), while the budding yeast S. cerevisiae contains a single ortholog, MOH1, previously linked to stress responses but with an unclear cellular role. Here, we investigated the function of MOH1 in S. cerevisiae. MOH1 deletion resulted in stress-specific phenotypes, including increased sensitivity to sodium azide and sulfuric acid, but enhanced resistance to hydrogen peroxide and acetic acid. Moh1 protein levels were dynamically regulated, decreasing upon hydrogen peroxide treatment and increasing in response to sulfuric acid. Morphological analyses including SEM revealed that moh1 Δ cells are rounder, form aggregates, and exhibit altered surface architecture independently of stress. RNA profiling and FTIR spectroscopy uncovered transcriptional reprogramming and metabolic remodeling, including alterations in lipid, protein, and cell wall polysaccharide levels and composition. Functional analyses showed that increased resistance to hydrogen peroxide is not due to altered mitochondrial ROS production but rather to reduced intracellular ROS accumulation. This effect is attributed to decreased cellular uptake resulting from altered permeability, supported by resistance to Congo red and sensitivity to SDS, consistent with cell envelope remodeling. Collectively, our findings identify Moh1 as a regulatory factor linking gene expression to metabolism and cellular architecture, thereby influencing cell envelope permeability and conferring selective stress resistance in S. cerevisiae.
Ubiquitously expressed Prohibitin-1 (PHB1) and Prohibitin-2 (PHB2) serve pleiotropic functions in cellular processes, including signal transduction, mitochondrial metabolism and dynamics, and lipid raft formation. Located in phospholipid-rich subcellular sites such as the plasma membrane and mitochondrial inner membrane, PHB1 and PHB2 are emerging as important host targets for bacteria and viruses, influencing infection and host responses by these microbes. Here, we present the current understanding of PHB1 and PHB2 in bacterial and viral infection based on the cellular localization of PHBs at the plasma membrane, mitochondria, and cytoplasm. We also discuss the potential of targeting PHBs as therapeutics for bacterial and viral infections.
Klebsiella pneumoniae (Kp) is a Gram-negative bacillus responsible for approximately 10% of nosocomial bacterial infections and one-third of Gram-negative bacterial infections in hospitalized patients. The rise of multidrug-resistant and hypervirulent strains makes it a significant public health issue. This study characterized carbapenem-resistant Kp (CR-Kp) strains isolated at the Hospital Clínico Universidad de Chile (HCUCH) 2021-2022 and explored associations with clinical characteristics. 45 CR-Kp strains from 29 patients in critical care units were analyzed. Mass spectrometry was used for species identification, and antimicrobial susceptibility was assessed by Kirby-Bauer disk diffusion. Clonality was determined using pulsed-field gel electrophoresis (PFGE), and multiplex PCR detected resistance and virulence genes. Clonal strains underwent whole-genome sequencing. PCR revealed the high prevalence of carbapenemase genes and extended-spectrum β -lactamases. PFGE identified nine clones, corresponding to sequence types ST25, ST45, ST307, and ST1161. Frequent virulence factors included siderophores and adhesins, while the capsular serotype K2 was present in 44% of isolates. No classical hypervirulence markers were detected. The presence of blaKPC correlated with more extended hospitalization. These findings reveal convergence between multidrug resistance and adaptive virulence traits rather than classical hypervirulence, highlighting evolving pathogenic strategies in high-risk CR-Kp clones circulating in Chile and emphasizing the need for enhanced molecular surveillance and infection protocols in critical care settings.
Clinical studies have uncovered associations between malocclusions and bacteria-related oral diseases. However, which malocclusion drives alternations in the oral microbiome remains unclear. Here, we identified occlusal type (OT, a major malocclusion classification parameter) as a key host structural regulator of the oral microbiome composition and metabolite profiles in adolescents. Regarding microbial composition: Prevotella and Veillonella species enriched in the OT-I group, whereas Neisseria and Haemophilus species predominated in the OT-II group. These differential distributions and unique microbial associations contributed to the formation of two distinct oral microbiome clusters (“stomatotypes”). In terms of gene functions, the OT-II group exhibited enrichment in “Environmental information processing” (EIP) pathways, “Human Diseases” (HD) pathways, and virulence-associated genes including relA and cpsB/cdsA. Significant differences in metabolite profiles were also observed between groups. Multi-omics analysis revealed positive intra-group associations and negative associations between-groups among representative oral microbes, functional pathways, and metabolites, with specific dipeptides identified as potential key microbe-modulated metabolites. Our results revealed the pivotal role of OT in shaping the variations of the oral microbiome and metabolite, offering novel insights into how host anatomical structure influences oral microecology.
Outer membrane vesicles (OMVs) have been increasingly recognized as common mediators of bacterial physiology in Gram-negative bacteria, including Vibrio species. The degree and function of OMV production can differ among strains and even within a single species. The secretion of OMVs is a prevalent trait among many Vibrio species, particularly in pathogenic organisms such as Vibrio cholerae, Vibrio vulnificus, and Vibrio parahaemolyticus. The OMVs released by these organisms are often associated with infection, transport of virulence factors into host cells, defense against stress, biofilm formation, flagella rotation, transportation of active enzymes, signaling molecules in the surrounding environment, and facilitating bacterial translocation. All of these are advantageous to the bacteria. These OMVs also possess immunogenic properties that regulate the innate and adaptive immune responses, which are beneficial to host cells. Few species, such as Vibrio ordalii, Vibrio coralliilyticus, Vibrio natriegens Vibrio alginolyticus, and Vibrio europaeus, have been recently studied for the first time that secrete OMVs; future research is necessary to determine any other activities that these vesicles may possess beyond those that are now documented.
Sugar-induced cell death (SICD) is a phenomenon observed in Saccharomyces cerevisiae whereby cells rapidly lose viability in glucose-only solutions. One theory suggests that SICD occurs due to an imbalance in nitrogen and carbon, however, limited studies are available to support this. When stationary phase cells are transferred to glucose-only solutions, cell death resembles that of apoptosis, while exponential phase cells show hallmarks of primary necrosis. Apoptosis in stationary phase cells is independent of the yeast metacaspase, YCA1, however, it remains unknown if SICD occurs through a caspase-independent pathway. Using stationary phase S. cerevisiae BY4741, we showed that SICD can be induced to the same degree with 10 mM or 110 mM glucose. Interestingly, SICD induced by 10 mM of glucose can be protected by supplementation with low concentrations of highly preferred organic nitrogen sources, namely glutamate, glutamine, and arginine, as well as high concentrations of non-preferred organic nitrogen sources. Additionally, cell death can be rescued by deletion of YCA1 and genes involved in caspase-independent apoptosis—STE20, NMA111, AIF1, or NUC1. On the other hand, when S. cerevisiae BY4741 is challenged with 110 mM glucose, SICD can only be rescued by supplementation with the same preferred organic nitrogen sources or deletion of AIF1 or NMA111. In all cases, protection is associated with a decrease in intracellular ROS and preservation of membrane integrity. Taken together, 110 mM glucose results in a catastrophic cell death phenotype that is more difficult to rescue, and nuclear localization of Aif1p and Nma111p is important for cell death in response to glucose.
The twin-arginine translocation (Tat) system is the only general pathway for the transport of folded proteins across energized biological membranes. It is found in the bacterial or archaeal cytoplasmic membrane, the plant thylakoid membrane or the inner membrane of plant mitochondria. The biological importance of this translocation system can be exemplified by the fact that all bacterial or plant photosynthesis and photosynthetic oxygen evolution on earth requires this system. Despite many biochemical and biophysical studies, the Tat mechanism has been puzzling since the system was discovered in the 1990ies. Important characteristics of the Tat system could not be explained, and also recent high-resolution structures of the Tat system’s core with bound substrate has not led to a general transport mechanism yet. In this integrative review, we attempted to answer the key open questions relevant to the Tat mechanism and thereby developed an in its molecular detail new comprehensive explanation of how folded proteins are translocated across membranes by the Tat system.
Chronic Obstructive Pulmonary Disease (COPD) is a progressive respiratory disease with high morbidity and mortality. Existing treatment methods are difficult to effectively curb disease progression, highlighting the urgency to explore new pathogenesis mechanisms and therapeutic targets. With the development of microbiomics, the proposal of the “gut-lung axis” concept has provided a brand-new perspective for understanding the pathological mechanisms of COPD, revealing that the gut and lungs maintain a close connection through pathways such as immune regulation and metabolic interaction. This article systematically elaborates on the association between gut microbiota and COPD: First, it deeply analyzes the pathological interaction between the gut and lungs from the perspective of the gut-lung axis. On this basis, it examines the characteristic changes in gut microbiota and their metabolites in COPD patients, explores the key influencing factors driving such microbiota dysbiosis, and further systematically explains the core mechanisms by which gut microbiota contribute to the occurrence and progression of COPD. Finally, it focuses on strategies for the prevention and treatment of COPD based on gut microbiota regulation, and prospects their clinical application potential. The purpose of this article is to provide new ideas and directions for the basic research and clinical practice of COPD by comprehensively sorting out the association between gut microbiota and COPD, thereby helping to improve the current status of COPD prevention and treatment.
The Juvenile form of Batten disease is a neurodegenerative disease with symptoms starting in the first decade and ending in death in the third decade of life. The gene defective in this form of Batten disease, CLN3, is conserved in eukaryotes, suggesting that the gene product serves a basic function in the cell, though the function is unknown. We have investigated the expression and regulation of the yeast homolog BTN1. Reanalysis of publicly available gene expression data suggests that transcription of BTN1 increases in response to oxidative stress, treatment with rapamycin or arsenate, amino acid starvation, and sporulation conditions. Similar to GCN4, there are upstream open reading frames (uORF) in front of BTN1, suggesting translational regulation. We developed reporter strains in which the HIS3 open reading frame replaced that of the BTN1 gene, with and without the uORFs. These reporters show that one or more of the uORFs decrease the expression of the HIS3 reporter. When expressed in the reporter strain using a high copy vector, GCN3, tRNA Arg , and tRNA Leu , increase expression, suggesting the involvement of the TORC1 pathway. BIT61 abuts BTN1 but is encoded on the opposite strand; 3' RACE analysis indicates that the mRNA of BIT61 overlaps with that of BTN1. BIT61 is involved in the TORC2 pathway, which interacts with the TORC1 pathway, suggesting a possible cis-acting mechanism of co-regulation. Lastly, we demonstrate that a yeast strain with a null mutation in BTN1 is sensitive to selective amino acid starvation, further supporting the association of BTN1 with TORC1.
Bacteria are important agents in the biodeterioration of cultural heritage objects, including historical manuscripts. Characterizing bacterial communities and generating robust microbiological data has therefore become crucial for conservation and restoration strategies. In this study, we investigated the bacterial communities associated with biodeterioration in six historical manuscripts using both culture-dependent and culture-independent (Illumina MiSeq) approaches. Culture-dependent methods yielded only 16 viable and culturable isolates, highlighting the limitations of traditional techniques. In contrast, metagenomic analysis revealed a far richer and more diverse bacterial community, capturing both living and non-living microbial traces accumulated over centuries. Bacterial genera with known cellulolytic and/or proteolytic activities, such as Bacillus, Stenotrophomonas, Pseudomonas and Acinetobacter, were identified as part of a core microbiome commonly associated with paper deterioration. High abundances of gut-associated bacteria (Prevotella, Faecalibacterium, Bacteroides, Porphyromonas) and human-related taxa (Staphylococcus, Streptococcus, Cutibacterium) indicated extensive historical human handling. A notable finding was the detection of Pseudonocardia broussonetiae, an endophytic bacterium associated with paper mulberry (Broussonetia papyrifera), suggesting the possible use of this plant as a papermaking material in one manuscript. This represents an important contribution to understanding Islamic paper production. Overall, our results demonstrate that effective conservation strategies require a detailed understanding of each manuscript’s microbial ecology, together with evidence of past environmental conditions, handling history, and production materials.
Vibrio natriegens, the fastest growing non-pathogenic microorganism known to date, has emerged as a highly promising chassis strain for synthetic biology and biotechnology applications. This study analysed the make-up and regulation of the biosynthetic pathway for L-lysine and related L-aspartate family amino acids (AFAAs) in V. natriegens DSM759 to provide a comprehensive basis for future metabolic engineering endeavours aiming at developing this strain into an amino acid overproducer. The compilation of automatically annotated genome sequencing data revealed the presence of gene duplicates encoding putative isozymes for multiple enzymatic reactions within these pathways. The physiological role of these isozymes was analysed via growth phenotyping of corresponding gene deletion mutants as well as enzymatic assays. We verified the presence of a previously unknown mono-functional aspartate kinase isozyme, here termed Vn.LysC2, which was shown to be insensitive to allosteric inhibition by any AFAA. In addition, functional duplicates of L-aspartate semialdehyde dehydrogenase and dihydrodipicolinate synthase enzymes were identified. RNA sequencing experiments were used to elucidate the transcriptional regulation mediated by AFAAs on both their corresponding biosynthetic pathways as well as on the global metabolism. The presence of L-lysine, L-threonine, L-isoleucine and L-methionine resulted in the transcriptional repression of their respective biosynthetic pathways. A global analysis of the transcriptional response revealed that the transcriptional response to L-lysine and L-isoleucine was characterised by a high degree of specificity (four and seven differentially expressed genes, respectively), while L-methionine and L-threonine supplementation affected the expression of a larger number of genes (37 and 60 differentially expressed genes).
Protein arginine methyltransferases (PRMTs) catalyse the transference of methyl groups from S-adenosylmethionine to arginine residues in substrate proteins, a post-translational modification widespread among eukaryotes. The change in size and hydrophobicity of the methylated arginine residue impacts on how a protein interacts with other macromolecules and affects several cellular processes, including intracellular signaling, DNA replication and repair, and control of gene expression. As a result, PRMTs became attractive targets for chemotherapy, and several PRMT inhibitors are going through clinical trials for cancer treatment. In protozoan parasites, PRMTs play fundamental roles during development, stage differentiation and infection processes. We here review the activity and the relevance of PRMTs for the survival of pathogenic kinetoplastids, apicomplexans and amoebas, highlight differences observed between PRMTs expressed in these organisms and their mammalian orthologues, and suggest that these enzymes can be exploited to combat parasitic infections. We propose that the arsenal of inhibitors developed to target mammalian PRMTs could be reassigned to allow the identification of new scaffolds to be explored as antiparasitic agents, either as sole chemotherapy or by improving the effectiveness of current antiparasitic drugs.
Capsular polysaccharides (CPS) are key virulence factors in Klebsiella pneumoniae and are closely associated with the K1 and K2 hypervirulent serotypes. Herein, we demonstrate that introducing nonspecific RNA (sgRNA) into K. pneumoniae ATCC43816 (Kp-pET-sgRNA), a K2 serotype strain classified as hypervirulent (hvKp), results in marked capsule loss and reduced hypermucoviscosity. Capsule loss and reduced hypermucoviscosity in Kp-pET-sgRNA were confirmed by comparison with the wild-type strain (Kp-WT) using transmission electron microscopy, hypermucoviscosity assays, and string tests. Mechanistically, we found that overexpression of sgRNA by introducing the pET-sgRNA plasmid led to gene deletion in the rmpADC operon, a key virulence determinant located on mobile chromosomal elements. Additionally, the mRNA expression of manC, which are chromosomal cps-related genes, was significantly repressed. In contrast, introduction of pET-sgRNA did not alter the mRNA expression of galF or wzi. The results revealed that capsule loss and reduced hypermucoviscosity in Kp-pET-sgRNA resulted from synergistic downregulation of both the rmpADC operon and manC. Loss of capsule synthesis and reduction of hypermucoviscosity in K. pneumoniae caused by sgRNA overexpression significantly decreased resistance to phagocytosis by macrophages but did not influence susceptibility to meropenem or colistin. The findings reveal an unexpected consequence of plasmid-mediated sgRNA introduction, where overexpression of sgRNA abolishes phagocytic resistance by disrupting capsule biosynthesis and reducing hypermucoviscosity in K. pneumoniae. This study highlights a promising strategy for disarming hypervirulent K. pneumoniae by directly targeting its key virulence factors and provides novel insights into antibacterial therapeutic approaches against this clinically significant pathogen.
Fungi were among the first eukaryotes to transition from aquatic to terrestrial life, developing multicellular hyphae, polar growth, and expanded secretomes for nutrient processing, defense, and symbiosis. We present a reliable method for purifying and characterizing extracellular vesicles (EVs) from Aspergillus nidulans and demonstrate that the induction of xylanase C is associated with increased EV release and EV-associated enzymatic activity. Using a mCherry reporter replacing xylanase C, we generalized this effect, showing that reporter induction increases EV production and reporter loading into EVs. This phenomenon primarily depends on the signal peptide (SP), suggesting that the induction of endoplasmic reticulum (ER)- trafficked proteins has a pronounced effect on EV production and cargo loading. We speculate that EV biogenesis may originate at the ER, where ER-translated proteins could be selectively loaded into vesicles and subsequently trafficked directly to the plasma membrane or through multivesicular bodies (MVBs). EV secretion is minimal in the first 24-48 hours but increases later in growth, coinciding with biofilm formation. This timing allows A. nidulans to modify the secretome, adapting it to new nutrient sources.
Autophagy contributes to cellular homeostasis by degrading and recycling intracellular components, especially under nutrient-limited conditions. While autophagy is well characterized under acute starvation in synthetic media in Saccharomyces cerevisiae, its regulation during the stationary phase of prolonged growth in nutrient-rich complex media, when cells experience gradual metabolic shifts and sustained stress, remains poorly understood. In this study, we identified Sir2, an NAD+-dependent histone deacetylase, as a key suppressor of autophagy during the stationary phase in YPD complex medium. Using GFP-Atg8 processing as a readout of autophagic flux, we demonstrated that SIR2 deletion led to sustained autophagy activation. Notably, Sir2 selectively inhibited mitophagy, pexophagy, and the Cvt pathway, while non-selective autophagy remained largely unaffected. Transcriptomic analysis revealed that Sir2 facilitates a coordinated entry into quiescence, in part by regulating ribosome biogenesis and nutrient-responsive pathways during the stationary phase. Mechanistically, Sir2 stabilized Ume6, a repressor of ATG8 transcription, thereby limiting autophagic activity. Deletion of SIR2 drastically increased the phosphorylation and stabilization of the mitochondrial receptor Atg32 during the stationary phase, leading to enhanced mitophagy. Additionally, we found that ROS generated by mitophagy enhanced autophagy through a positive feedback loop. Collectively, our findings establish Sir2 as a previously unrecognized regulator of selective autophagy during the stationary phase in complex medium and highlight how cells dynamically control organelle degradation to maintain viability under extended metabolic stress.
Cells typically balance growth with stress responses - growing rapidly in low stress conditions and halting growth to defend against stress or to repair stress-induced damage. While numerous genome-wide screens have identified mutants resistant to oxidative stress, these have largely relied on static, end-point measurements. Here, we take a dynamic, time-resolved approach to uncover how fission yeast, Schizosaccharomyces pombe, adapts to oxidative stress over time. We have tracked the growth of 3,420 deletion mutants across nine time points spanning four days on both nutrient-rich solid media and media containing oxidative stress induced by hydrogen peroxide. This kinetic strategy revealed not just resistant or sensitive mutants. It allowed clustering of growth patterns across time and uncovered mutants that are capable of transiently uncoupling growth from stress response. Hydrogen peroxide induced a dose-dependent delay in colony expansion in most deletion strains, yet 15 mutants consistently maintained robust growth. These belong to different functional categories, highlighting diverse potential mechanisms ranging from altered DNA damage checkpoints to metabolic rewiring and growth regulation. By capturing dynamic trajectories rather than static outcomes, this study exposes hidden layers of growth under oxidative stress and identifies new genetic determinants of cellular resilience in fission yeast.
