ACS Chemical Biology最新文献

{"title":"Biochemical Investigation of PumG, a Key Pyridoxal 5′-Phosphate (PLP)-Dependent Aminotransferase Involved in Pseudouridimycin Biosynthesis","authors":"Krushnamurthy Pattanayakanahalli Henjarappa,&nbsp;Mahima Ganguly,&nbsp;Aritra Mukherjee,&nbsp;Pooja Bhattoo,&nbsp;Yashwanth Naik,&nbsp;Krishna Mhaske,&nbsp;Dhananjaya Giriraju,&nbsp;Simita Das,&nbsp;Rishikesh Narayan,&nbsp;Shivajirao L. Gholap,&nbsp;Supriya Rej,&nbsp;Suvamay Jana,&nbsp;Nilkamal Mahanta","doi":"10.1021/acschembio.6c00500","DOIUrl":"10.1021/acschembio.6c00500","url":null,"abstract":"<p>Pyridoxal 5′-phosphate (PLP)-dependent aminotransferases play a central role in nucleoside antibiotic biosynthesis, where they catalyze the conversion of ribose-derived aldehyde intermediates into aminonucleosides by introducing critical amino functional groups. Despite their widespread occurrence in the biosynthetic pathways of several therapeutically important nucleoside antibiotics, the detailed biochemical and mechanistic characterization of these enzymes remains elusive. In this study, we present a comprehensive <em>in vitro</em> reconstitution, structural modeling, and molecular dynamics (MD) simulation-based biochemical and mechanistic investigation of a key aminotransferase, PumG from <em>Streptomyces rimosus</em> (SrPumG), which plays a critical role in the biosynthesis of the potent bacterial RNA polymerase inhibitor, Pseudouridimycin (PUM). Our size-exclusion chromatography and native PAGE analysis, in conjunction with spectroscopic and modeling studies, showed that SrPumG is a stable homodimer that uses a highly conjugated form of the PLP cofactor, along with suitable amino donors, such as L-Arg, to convert pseudouridine aldehyde (PUA) to amino pseudouridine (APU). The structure-based three-dimensional modeling and MD simulation of SrPumG, coupled with site-directed mutagenesis, further confirmed that, in addition to the catalytic residue Lys289, several active-site residues from both protomers facilitate the binding and stabilization of the PLP/PMP cofactor and the PUA substrate via electrostatic/hydrogen-bonding interactions and aromatic stacking. In addition, our studies uncovered key residues that stabilize the dimer interface via hydrophobic and π–π interactions. One notable finding from our study is that SrPumG does not discriminate between <em>C</em>- and <em>N</em>-nucleoside substrates and exhibits a broader substrate scope, which further confirms the gatekeeper role of the preceding oxidoreductase enzyme, PumI, in this pathway. Together, these findings reveal that a distinct highly conjugated form of PLP and an intricate dimer interface architecture, which is essential for substrate/cofactor binding and protein stability, govern SrPumG catalysis. This work provides new insights into aminotransferases in nucleoside biosynthesis and establishes a foundation for engineering the PUM pathway to design new antibacterial derivatives.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2630–2649"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149007661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ebselen Inactivates Essential Mycoplasmopsis bovis Proteins","authors":"Poonam Dhindwal,&nbsp;Nishad Thamban Chandrika,&nbsp;Yunyun Di,&nbsp;Sylvie Garneau-Tsodikova,&nbsp;Antonio Ruzzini","doi":"10.1021/acschembio.6c00562","DOIUrl":"10.1021/acschembio.6c00562","url":null,"abstract":"<p><em>Mycoplasmopsis bovis</em> contributes to a variety of infectious diseases in cattle that limit meat and milk production, motivating significant antimicrobial use to prevent animal and economic loss. Currently approved antimicrobial treatments for <em><em>M. bovis</em></em> are ineffective or becoming so due to adaptive antimicrobial resistance (AMR). Thus, to discover alternatives to these therapies, we screened a drug repurposing library of ∼2800 compounds as antibiotics, resulting in the prioritization of ebselen (Eb) for study. The ability of Eb to prevent <em>M. bovis</em> growth could be ascribed to the covalent modification and inhibition of key metabolic targets, including lactate dehydrogenase (Ldh), enolase (Eno) and elongation factor G. The identification, validation, and consequences of covalent modifications by Eb at specific sites in Ldh and Eno were determined using mass spectrometry and steady-state kinetic experiments. Specifically, non-catalytic cysteine residues that are modified by Eb – Ldh Cys124 and Eno Cys216 – result in enzyme inactivation with low micromolar IC₅₀ values. The poorly conserved Eb modification sites in <em>M. bovis</em> offer an opportunity for future narrow-spectrum therapeutic development for this major agricultural pathogen.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2695–2701"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149100966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An Automated Scalable Chemoproteomics Workflow that Recovers Known Serine Hydrolase Target Profiles of Clinically Used Drugs","authors":"Franciscus H.G. ter Brake,&nbsp;Berend Gagestein,&nbsp;Mario van der Stelt,&nbsp;Antonius P.A. Janssen","doi":"10.1021/acschembio.6c00557","DOIUrl":"10.1021/acschembio.6c00557","url":null,"abstract":"<p>Activity-based protein profiling has established itself as a powerful technology with demonstrated use in on- and off-target profiling. However, determining the off-target profile of many drugs using the unbiased mass spectrometry-based chemoproteomics approach remains time- and cost-intensive. In this study, we implemented an SP3-based, high-throughput, plate-based chemoproteomic workflow compatible with liquid-handling robotics. We then profiled a library of 40 marketed drugs targeting the serine hydrolases or containing potentially reactive electrophiles against the human serine hydrolase proteome in HT29 cell lysate. We included two additional well-studied literature compounds as controls to benchmark our workflow. Our results validated 16 out of 20 intended human targets and found 5 previously described off-targets for the compounds tested. Additionally, we discovered six unknown off-targets that could warrant future follow-up research. In all, our approach demonstrates how automated chemoproteomics can be used to accelerate the identification of clinically relevant off-targets, providing a scalable blueprint for rational drug repurposing.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2683–2694"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148917753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Minimal Functional Scaffolds of the Streptococcus sinensis Competence-Stimulating Peptide Define the Balance between Receptor Promiscuity and Signaling Specificity","authors":"Clay P. Renshaw,&nbsp;Angela Wang,&nbsp;Christopher R. Yeh,&nbsp;Aspen C. Easter,&nbsp;Umme N. Simanto,&nbsp;Yftah Tal-Gan","doi":"10.1021/acschembio.6c00715","DOIUrl":"10.1021/acschembio.6c00715","url":null,"abstract":"<p>Quorum sensing (QS) enables bacteria to coordinate collective behaviors through the detection of secreted signaling molecules and represents an attractive target for anti-virulence therapeutic development. In <em>Streptococcus sinensis</em>, an emerging opportunistic pathogen associated with infective endocarditis, QS is mediated by a competence-stimulating peptide (CSP) that activates the membrane-bound histidine kinase receptor ComD. Although previous studies demonstrated that the <em>S. sinensis</em> CSP is unusually tolerant to single-amino acid substitutions, the minimum structural requirements for productive receptor activation remained unknown. Here, we systematically define the molecular determinants of <em>S. sinensis</em> CSP activity through peptide truncation analysis, reverse alanine scanning, and inter-species crosstalk screening. Truncation studies identified the <em>C</em>-terminal Lys as dispensable for activity while highlighting the critical role of the <em>N</em>-terminus in receptor engagement. Reverse alanine scanning revealed multiple minimal functional scaffolds that retained native levels of QS activation despite replacement of nearly one-third of the peptide sequence with alanine, demonstrating that productive signaling requires only a limited subset of native residues. Consistent with these findings, screening of CSPs from diverse <em>Streptococcus</em> species revealed that only a small subset activated the <em>S. sinensis</em> ComD receptor, and only with substantially reduced potency relative to the native CSP, indicating that receptor permissiveness remains constrained by specific sequence requirements. Finally, incorporation of a D1A substitution predictably converted agonists, including minimal functional scaffolds, into competitive antagonists while preserving their relative potency trends. The most potent antagonist also demonstrated phenotypic suppression of natural competence in <em>S. sinensis</em>, confirming functional inhibition of QS-controlled behavior. Together, these findings define the minimal structural features required for <em>S. sinensis</em> QS activation, provide new insight into how receptor promiscuity is balanced with signaling specificity, and establish a foundation for the rational design of peptide-based QS modulators.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2811–2818"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149100915","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultramutagenesis through Combined Msh2 Depletion and Proofreading-Deficient DNA Polymerase ε Expression Enhances Small-Molecule Target Identification in Forward Genetic Screens","authors":"Alexander-Hoi Nguyen,&nbsp;Vicky Li,&nbsp;Wei-Min Chen,&nbsp;Jacob Kimberg,&nbsp;Anthony J. Davis,&nbsp;David G. McFadden,&nbsp;Juan Manuel Povedano","doi":"10.1021/acschembio.6c00605","DOIUrl":"10.1021/acschembio.6c00605","url":null,"abstract":"<p>DNA mismatch repair (MMR) deficiency has been widely utilized in forward genetic screens to identify drug resistance mutations and elucidate the mechanisms of action of cytotoxic small molecules. However, MMR deficiency generates a characteristic mutational signature enriched for C &gt; T transitions that precludes saturation of mutagenesis. We hypothesized that expression of proofreading-deficient DNA polymerase epsilon, as has been observed in patients with biallelic MMR deficiency, would enhance mutagenesis and enable identification of new resistance mutations and cytotoxin mechanisms. Here, we combine auxin-inducible degradation of Msh2 with doxycycline-inducible expression of the proofreading-deficient <em>Pol</em>ε^P286R mutant in murine small cell lung cancer (SCLC) cells. Simultaneous MMR deficiency and mutant <em>Pol</em>ε expression markedly increased the emergence of bortezomib-resistant clones relative to either perturbation alone, and targeted sequencing of <em>Psmb5</em> in bortezomib-resistant clones identified recurrent mutations previously reported in resistance screens using MMR-deficient cells and chemical mutagenesis. We next applied this platform to investigate resistance to lurbinectedin, a clinically relevant therapy for SCLC. Whole-exome sequencing revealed recurrent loss-of-function alterations in nucleotide excision repair (NER) genes, most notably <em>Ercc5</em> and <em>Ercc4</em>, implicating NER deficiency as a major mechanism of lurbinectedin resistance and supporting a central role for NER in mediating lurbinectedin-induced cytotoxicity. Collectively, these findings establish inducible ultramutagenesis as a powerful and versatile platform for the unbiased discovery of drug resistance mechanisms and therapeutic vulnerabilities.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2755–2764"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148928570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering a Ribozyme with Aminoacyl-tRNA Synthetase Activity","authors":"Rui Gan,&nbsp;Ji Chen,&nbsp;Gerald Manuel,&nbsp;Hillary Andaluz,&nbsp;W. Andy Tao,&nbsp;Robert M. Corn,&nbsp;Donald H. Burke,&nbsp;Andrej Luptak,&nbsp;Barbara L. Golden","doi":"10.1021/acschembio.6c00709","DOIUrl":"10.1021/acschembio.6c00709","url":null,"abstract":"<p>A ribozyme that can charge a tRNA with amino acids and discriminate between cognate and non-cognate tRNAs is of interest because an RNA with this ability may have been critical for translation in the transition from the RNA world. In addition, a fully optimized ribozyme could provide a tool for incorporating non-canonical amino acids for biotechnology applications. Here, we rationally engineer a ribozyme by fusing a tRNA binding module derived from a T-box riboswitch with a catalytic module (a flexizyme) to generate a ribozyme that can aminoacylate a target tRNA. We demonstrate that this ribozyme can be readily redesigned to alter tRNA specificity. This ribozyme is compatible with an in vitro translation system and could be used to recode a protein sequence to site-specifically incorporate a non-canonical amino acid.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2800–2810"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149056660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"On the Inhibition of Lipid Peroxidation and Ferroptosis by Fatty Acids","authors":"Melodie Mallais,&nbsp;Krystina Szylo,&nbsp;Leslie Magtanong,&nbsp;Mikhail S. Shchepinov,&nbsp;Scott J. Dixon,&nbsp;Derek A. Pratt","doi":"10.1021/acschembio.6c00583","DOIUrl":"10.1021/acschembio.6c00583","url":null,"abstract":"<p>Most inhibitors of lipid peroxidation (LPO) and associated ferroptosis are small molecules that trap LPO-propagating radicals. Among the handful of other inhibitors are select fatty acids: monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids substituted with deuterium atoms at their reactive bis-allylic positions (D-PUFAs), which render them significantly less reactive to LPO-propagating radicals. To probe whether the simple replacement of oxidizable PUFAs with non-oxidizable FAs is a general strategy for ferroptosis suppression, we prepared derivatives of representative PUFAs—linoleic acid (LA) and arachidonic acid (AA)—with cyclopropane rings in place of their unsaturations (CP-PUFAs). Cyclopropanation was predicted to boost the strength of the neighboring C–H bonds by ∼20 kcal/mol and increase the barrier to reaction with peroxyl radicals by ∼10⁴-fold while preserving their <em>cis</em> geometry. CP-PUFAs suppressed ferroptosis induced by erastin2 in HT-1080 cells and RSL3 in HEK-293 cells, similarly to D-PUFAs and MUFAs. Palmitate, a representative endogenous saturated FA, did not suppress ferroptosis. Whereas <em>d</em>₆-AA was more effective than <em>d</em>₂-LA, the opposite was true of the CP-PUFAs, with CP₄-AA possessing only modest activity while CP₂-LA was comparable to the D-PUFAs. Lipidomics provides evidence for more extensive lipid remodeling upon treatment with CP₂-LA relative to CP₄-AA, with PUFAs being enriched in triacylglycerols at the expense of the diacylglycerols used for phospholipid synthesis. Overall, these results suggest that replacement of oxidizable PUFAs with non-oxidizable FAs is a general strategy to suppress ferroptosis—provided that the non-oxidizable FA can be utilized by the biosynthetic machinery and is not lipotoxic at concentrations necessary for protection.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2702–2711"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149072796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of m6A Readers for Enhanced Recognition In Vitro via Genetic Code Expansion","authors":"Michael Addo,&nbsp;Elise Ficaretta,&nbsp;Yibo Chang,&nbsp;Weiqi Qiu,&nbsp;Chong Teng,&nbsp;Junwei Lucas Bao,&nbsp;Abhishek Chatterjee,&nbsp;Huiqing Zhou","doi":"10.1021/acschembio.6c00476","DOIUrl":"10.1021/acschembio.6c00476","url":null,"abstract":"<p>The YTH domain was recently demonstrated as a promising tool for recognizing and regulating m⁶A-modified RNAs. However, the deficiency in binding affinity (micromolar range) and selectivity (off-target binding) for m⁶A of the wild-type YTH domain limit its sensitivity and accuracy for m⁶A recognition. Engineering strategies for improving the affinity and selectivity of such epitranscriptomic readers remain lacking. Here, we demonstrate that site-specific introduction of noncanonical tryptophan derivatives within the aromatic m⁶A-binding pocket of the YTH domain modulates the binding affinity and selectivity. Specifically, we report two YTH variants, with 5-bromotryptophan and 5-hydroxytryptophan incorporated at the W465 of the YTH domain of YTHDF1, which show improved binding affinity and selectivity against m⁶A-modified RNAs. DFT-based energy decomposition analysis indicates that the binding affinity can be modulated through the substitution group’s effect on the dispersion force in the van der Waals recognition complex, suggesting a critical role of the CH–π interaction in m⁶A-reader recognition. This work reports new reader domains with improved m⁶A recognition and demonstrates the potential of m⁶A reader engineering via genetic code expansion.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 10","pages":"2611–2621"},"PeriodicalIF":3.6,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148917748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Benzo[c]acridine Dimer Cyclization as a Route toward a Small Bicationic Heteroaromatic Compound with Unprecedented G-Quadruplex Stabilization Properties.","authors":"Stefanie Sandra Sandberg, Delphine Pitrat, Marine Labro, Sepideh Farmand Azadeh, Marco Deiana, Tangui Le Bahers, Sandrine Denis-Quanquin, Cyrille Monnereau, Natacha Gillet, Nasim Sabouri, Laure Guy","doi":"10.1021/acschembio.6c00586","DOIUrl":"https://doi.org/10.1021/acschembio.6c00586","url":null,"abstract":"<p><p>G-quadruplex (G4) DNA structures are key regulatory elements in genome stability and gene expression, with a pronounced role in cancer biology due to their selective formation in oncogenic genomic regions and abundance in tumor cells. Their structural uniqueness makes them compelling targets for therapeutic intervention. Thus, targeting G4s with small molecule ligands offers a promising approach for modulating gene expression and inducing replication stress, particularly in DNA damage response-deficient malignancies such as ovarian and breast cancers. However, the structural diversity of G4s has hindered the development of ligands capable of broad-spectrum, topology-independent stabilization. Here, we introduce a novel dimeric N-methylbenzo[c]acridinium derivative, engineered as a visible-light-absorbing cationic G4 ligand. Compared to previously reported diazonia derivatives, this compound demonstrates red-shifted optical properties, superior luminescence quantum efficiency, and outstanding G4-stabilizing activity. It exhibits high preferential binding for G4s over dsDNA, induces conformational transitions between hybrid and antiparallel G4s, stalls DNA replication, and disrupts DNA duplex formation. Notably, its broad, topology-independent recognition enables stabilization of a range of G4 targets. Crucially, it surpasses benchmark G4 ligands, including PhenDC3 and CX-5461, in both selectivity and/or efficacy. These results support the dimeric benzo[c]acridinium scaffold as a versatile platform for the development of G4-targeted therapeutic strategies.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149190212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pooled Screening of Cancer Cell Lines Identifies Vulnerability to Polyunsaturated Fatty-Acid-Induced Ferroptosis.","authors":"Zijian Pan, Jingrong Regina Han, Nathchar Naowarojna, Emily T Graham, Jennifer A Roth, Wenyu Wang, Guanglei Xie, Xi Wang, Haoxin Li, Yilong Zou","doi":"10.1021/acschembio.6c00581","DOIUrl":"https://doi.org/10.1021/acschembio.6c00581","url":null,"abstract":"<p><p>Polyunsaturated fatty acids (PUFAs) are essential nutrients supporting numerous cellular processes in normal and neoplastic cells, yet PUFA-containing phospholipids predispose cells to ferroptosis. How cancer cells from various lineages or genetic backgrounds proliferate in the presence of PUFA remains poorly understood. Here, by systematically profiling various fatty acids using a barcoded and pooled cancer cell line library, we show that a subset of cell lines is sensitive to PUFA treatment at concentrations comparable to physiological levels. We demonstrate that PUFA treatment alone is sufficient to induce ferroptosis in these cell lines that are also vulnerable to GPX4 inhibition. By correlating patterns of PUFA sensitivity with cancer cell line features including basal mRNA transcript levels, we show that PUFA sensitivity is positively associated with fatty acid binding protein 1 (FABP1) expression. Additionally, by profiling a diverse collection of fatty acids and analogs, we find that ferroptosis in cancer cells can be effectively induced by conjugated linolenic acids, in addition to PUFAs containing 1,4-pentadiene groups. Genome-wide CRISPR modifier screens in the PUFA-sensitive ES2 cells reveal a pro-ferroptosis pathway defined by enzymes that convert PUFAs to membrane-associated phospholipids, and an anti-ferroptosis pathway that segregates PUFAs by delivering them to lipid droplets. Together, we present the first comprehensive data set of PUFA-induced ferroptosis sensitivity in a collection of 554 cell lines from 19 primary lineages and suggest a potential therapeutic avenue to induce tumor cell ferroptosis using dietary metabolites.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149177945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}