EMBO Journal最新文献_第10页

RNA-binding proteins hnRNPM and ELAVL1 promote type-I interferon induction downstream of the nucleic acid sensors cGAS and RIG-I. rna结合蛋白hnRNPM和ELAVL1促进i型干扰素在核酸传感器cGAS和rig - 1下游的诱导。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2024-12-20 DOI: 10.1038/s44318-024-00331-x

Alexander Kirchhoff, Anna-Maria Herzner, Christian Urban, Antonio Piras, Robert Düster, Julia Mahlberg, Agathe Grünewald, Thais M Schlee-Guimarães, Katrin Ciupka, Petro Leka, Robert J Bootz, Christina Wallerath, Charlotte Hunkler, Ann Kristin de Regt, Beate M Kümmerer, Maria Hønholt Christensen, Florian I Schmidt, Min Ae Lee-Kirsch, Claudia Günther, Hiroki Kato, Eva Bartok, Gunther Hartmann, Matthias Geyer, Andreas Pichlmair, Martin Schlee

{"title":"RNA-binding proteins hnRNPM and ELAVL1 promote type-I interferon induction downstream of the nucleic acid sensors cGAS and RIG-I.","authors":"Alexander Kirchhoff, Anna-Maria Herzner, Christian Urban, Antonio Piras, Robert Düster, Julia Mahlberg, Agathe Grünewald, Thais M Schlee-Guimarães, Katrin Ciupka, Petro Leka, Robert J Bootz, Christina Wallerath, Charlotte Hunkler, Ann Kristin de Regt, Beate M Kümmerer, Maria Hønholt Christensen, Florian I Schmidt, Min Ae Lee-Kirsch, Claudia Günther, Hiroki Kato, Eva Bartok, Gunther Hartmann, Matthias Geyer, Andreas Pichlmair, Martin Schlee","doi":"10.1038/s44318-024-00331-x","DOIUrl":"10.1038/s44318-024-00331-x","url":null,"abstract":"The cytosolic nucleic acid sensors RIG-I and cGAS induce type-I interferon (IFN)-mediated immune responses to RNA and DNA viruses, respectively. So far no connection between the two cytosolic pathways upstream of IKK-like kinase activation has been investigated. Here, we identify heterogeneous nuclear ribonucleoprotein M (hnRNPM) as a positive regulator of IRF3 phosphorylation and type-I IFN induction downstream of both cGAS and RIG-I. Combining interactome analysis with genome editing, we further uncover the RNA-binding protein ELAV-like protein 1 (ELAVL1; also known as human antigen R, HuR) as an hnRNPM interactor. Depletion of hnRNPM or ELAVL1 impairs type-I IFN induction by herpes simplex virus 1 or Sendai virus. In addition, we show that hnRNPM and ELAVL1 interact with TANK-binding kinase 1, IκB kinase ε, IκB kinase β, and NF-κB p65. Our confocal microscopy experiments demonstrate cytosolic and perinuclear interactions between hnRNPM, ELAVL1, and TBK1. Furthermore, pharmacological inhibition of ELAVL1 strongly reduces cytokine release from type-I interferonopathy patient fibroblasts. The RNA-binding proteins hnRNPM and ELAVL1 are the first non-redundant regulators to bridge the cGAS/STING and RIG-I/MAVS pathways. Overall, our study characterizes the hnRNPM-ELAVL1 complex as a novel system promoting antiviral defense, pointing to a potential therapeutic target to reduce auto-inflammation in patients with type-I interferonopathies.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"824-853"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11791083/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142873199","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Partitioning of fatty acids between membrane and storage lipids controls ER membrane expansion. 脂肪酸在膜和储存脂质之间的分配控制内质网膜的扩张。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-03 DOI: 10.1038/s44318-024-00355-3

Pawel K Lysyganicz, Antonio D Barbosa, Shoily Khondker, Nicolas A Stewart, George M Carman, Phillip J Stansfeld, Marcus K Dymond, Symeon Siniossoglou

{"title":"Partitioning of fatty acids between membrane and storage lipids controls ER membrane expansion.","authors":"Pawel K Lysyganicz, Antonio D Barbosa, Shoily Khondker, Nicolas A Stewart, George M Carman, Phillip J Stansfeld, Marcus K Dymond, Symeon Siniossoglou","doi":"10.1038/s44318-024-00355-3","DOIUrl":"10.1038/s44318-024-00355-3","url":null,"abstract":"Biogenesis of membrane-bound organelles involves the synthesis, remodeling, and degradation of their constituent phospholipids. How these pathways regulate organelle size remains poorly understood. Here we demonstrate that a lipid-degradation pathway inhibits expansion of the endoplasmic reticulum (ER) membrane. Phospholipid diacylglycerol acyltransferases (PDATs) use endogenous phospholipids as fatty-acyl donors to generate triglyceride stored in lipid droplets. The significance of this non-canonical triglyceride biosynthesis pathway has remained elusive. We find that the activity of the yeast PDAT Lro1 is regulated by a membrane-proximal helical segment facing the luminal side of the ER bilayer. To reveal the biological roles of PDATs, we engineered an Lro1 variant with derepressed activity. We show that active Lro1 mediates retraction of ER membrane expansion driven by phospholipid synthesis. Furthermore, subcellular distribution and membrane turnover activity of Lro1 are controlled by diacylglycerol produced by the activity of Pah1, a conserved member of the lipin family. Collectively, our findings reveal a lipid-metabolic network that regulates endoplasmic reticulum biogenesis by converting phospholipids into storage lipids.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"781-800"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11790888/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Developmental beta-cell death orchestrates the islet's inflammatory milieu by regulating immune system crosstalk. 发育中的β细胞死亡通过调节免疫系统的串扰来协调胰岛的炎症环境。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-06 DOI: 10.1038/s44318-024-00332-w

Mohammad Nadeem Akhtar, Alisa Hnatiuk, Luis Delgadillo-Silva, Shirin Geravandi, Katrin Sameith, Susanne Reinhardt, Katja Bernhardt, Sumeet Pal Singh, Kathrin Maedler, Lutz Brusch, Nikolay Ninov

{"title":"Developmental beta-cell death orchestrates the islet's inflammatory milieu by regulating immune system crosstalk.","authors":"Mohammad Nadeem Akhtar, Alisa Hnatiuk, Luis Delgadillo-Silva, Shirin Geravandi, Katrin Sameith, Susanne Reinhardt, Katja Bernhardt, Sumeet Pal Singh, Kathrin Maedler, Lutz Brusch, Nikolay Ninov","doi":"10.1038/s44318-024-00332-w","DOIUrl":"10.1038/s44318-024-00332-w","url":null,"abstract":"While pancreatic beta-cell proliferation has been extensively studied, the role of cell death during islet development remains incompletely understood. Using a genetic model of caspase inhibition in beta cells coupled with mathematical modeling, we here discover an onset of beta-cell death in juvenile zebrafish, which regulates beta-cell mass. Histologically, this beta-cell death is underestimated due to phagocytosis by resident macrophages. To investigate beta-cell apoptosis at the molecular level, we implement a conditional model of beta-cell death linked to Ca2+ overload. Transcriptomic analysis reveals that metabolically-stressed beta cells follow paths to either de-differentiation or apoptosis. Beta cells destined to die activate inflammatory and immuno-regulatory pathways, suggesting that cell death regulates the crosstalk with immune cells. Consistently, inhibiting beta-cell death during development reduces pro-inflammatory resident macrophages and expands T-regulatory cells, the deficiency of which causes premature activation of NF-kB signaling in beta cells. Thus, developmental cell death not only shapes beta-cell mass but it also influences the islet's inflammatory milieu by shifting the immune-cell population towards pro-inflammatory.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":"44 4","pages":"1131-1153"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11833124/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143442566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

MTCH2 controls energy demand and expenditure to fuel anabolism during adipogenesis. 在脂肪形成过程中，MTCH2控制能量需求和消耗以促进合成代谢。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-03 DOI: 10.1038/s44318-024-00335-7

Sabita Chourasia, Christopher Petucci, Clarissa Shoffler, Dina Abbasian, Hu Wang, Xianlin Han, Ehud Sivan, Alexander Brandis, Tevie Mehlman, Sergey Malitsky, Maxim Itkin, Ayala Sharp, Ron Rotkopf, Bareket Dassa, Limor Regev, Yehudit Zaltsman, Atan Gross

{"title":"MTCH2 controls energy demand and expenditure to fuel anabolism during adipogenesis.","authors":"Sabita Chourasia, Christopher Petucci, Clarissa Shoffler, Dina Abbasian, Hu Wang, Xianlin Han, Ehud Sivan, Alexander Brandis, Tevie Mehlman, Sergey Malitsky, Maxim Itkin, Ayala Sharp, Ron Rotkopf, Bareket Dassa, Limor Regev, Yehudit Zaltsman, Atan Gross","doi":"10.1038/s44318-024-00335-7","DOIUrl":"10.1038/s44318-024-00335-7","url":null,"abstract":"Mitochondrial carrier homolog 2 (MTCH2) is a regulator of apoptosis, mitochondrial dynamics, and metabolism. Loss of MTCH2 results in mitochondrial fragmentation, an increase in whole-body energy utilization, and protection against diet-induced obesity. In this study, we used temporal metabolomics on HeLa cells to show that MTCH2 deletion results in a high ATP demand, an oxidized cellular environment, and elevated utilization of lipids, amino acids, and carbohydrates, accompanied by a decrease in several metabolites. Lipidomics analysis revealed a strategic adaptive reduction in membrane lipids and an increase in storage lipids in MTCH2 knockout cells. Importantly, MTCH2 knockout cells showed an increase in mitochondrial oxidative function, which may explain the higher energy demand. Interestingly, this imbalance in energy metabolism and reductive potential triggered by MTCH2-deletion prevents NIH3T3L1 preadipocytes from differentiating into mature adipocytes, an energy consuming reductive biosynthetic process. In summary, the loss of MTCH2 leads to increased mitochondrial oxidative activity and energy demand, creating a catabolic and oxidative environment that fails to fuel the anabolic processes required for lipid accumulation and adipocyte differentiation.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"1007-1038"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11832942/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RNA dysregulation in neurodegenerative diseases. 神经退行性疾病中的RNA失调。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-09 DOI: 10.1038/s44318-024-00352-6

Yini Li, Shuying Sun

引用次数: 0

Hypoxia promotes tumor immune evasion by suppressing MHC-I expression and antigen presentation. 缺氧通过抑制mhc - 1表达和抗原呈递促进肿瘤免疫逃避。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-03 DOI: 10.1038/s44318-024-00319-7

Hala Estephan, Arun Tailor, Robert Parker, McKenzie Kreamer, Ioanna Papandreou, Leticia Campo, Alistair Easton, Eui Jung Moon, Nicholas C Denko, Nicola Ternette, Ester M Hammond, Amato J Giaccia

{"title":"Hypoxia promotes tumor immune evasion by suppressing MHC-I expression and antigen presentation.","authors":"Hala Estephan, Arun Tailor, Robert Parker, McKenzie Kreamer, Ioanna Papandreou, Leticia Campo, Alistair Easton, Eui Jung Moon, Nicholas C Denko, Nicola Ternette, Ester M Hammond, Amato J Giaccia","doi":"10.1038/s44318-024-00319-7","DOIUrl":"10.1038/s44318-024-00319-7","url":null,"abstract":"Hypoxia is a common feature of solid tumors that has previously been linked to resistance to radiotherapy and chemotherapy, and more recently to immunotherapy. In particular, hypoxic tumors exclude T cells and inhibit their activity, suggesting that tumor cells acquire a mechanism to evade T-cell recognition and killing. Our analysis of hypoxic tumors indicates that hypoxia downregulates the expression of MHC class I and its bound peptides (i.e., the immunopeptidome). Hypoxia decreases MHC-I expression in an oxygen-dependent manner, via activation of autophagy through the PERK arm of the unfolded protein response. Using an immunopeptidomics-based LC-MS approach, we find a significant reduction of presented antigens under hypoxia. Inhibition of autophagy under hypoxia enhances antigen presentation. In experimental tumors, reducing mitochondrial metabolism through a respiratory complex-I inhibitor increases tumor oxygenation, as well as MHC-I levels and the immunopeptidome. These data explain the molecular basis of tumor immune evasion in hypoxic conditions, and have implications for future therapeutic interventions targeting hypoxia-induced alterations in antigen presentation.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"903-922"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11790895/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The integrated stress response drives MET oncogene overexpression in cancers. 综合应激反应驱动MET癌基因在癌症中的过度表达。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-07 DOI: 10.1038/s44318-024-00338-4

Marina Cerqua, Marco Foiani, Carla Boccaccio, Paolo M Comoglio, Dogus M Altintas

{"title":"The integrated stress response drives MET oncogene overexpression in cancers.","authors":"Marina Cerqua, Marco Foiani, Carla Boccaccio, Paolo M Comoglio, Dogus M Altintas","doi":"10.1038/s44318-024-00338-4","DOIUrl":"10.1038/s44318-024-00338-4","url":null,"abstract":"Cancer cells rely on invasive growth to survive in a hostile microenvironment; this growth is characterised by interconnected processes such as epithelial-to-mesenchymal transition and migration. A master regulator of these events is the MET oncogene, which is overexpressed in the majority of cancers; however, since mutations in the MET oncogene are seen only rarely in cancers and are relatively infrequent, the mechanisms that cause this widespread MET overexpression remain obscure. Here, we show that the 5' untranslated region (5'UTR) of MET mRNA harbours two functional stress-responsive elements, conferring translational regulation by the integrated stress response (ISR), regulated by phosphorylation of eukaryotic translation initiation factor 2 alpha (eIF2α) at serine 52. ISR activation by serum starvation, leucine deprivation, hypoxia, irradiation, thapsigargin or gemcitabine is followed by MET protein overexpression. We mechanistically link MET translation to the ISR by (i) mutation of the two uORFs within the MET 5'UTR, (ii) CRISPR/Cas9-mediated mutation of eIF2α (S52A), or (iii) the application of ISR pathway inhibitors. All of these interventions reduce stress-induced MET overexpression. Finally, we show that blocking stress-induced MET translation blunts MET-dependent invasive growth. These findings indicate that upregulation of the MET oncogene is a functional requirement linking integrated stress response to cancer progression.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"1107-1130"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11832788/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142958212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A nuclear protein quality control system for elimination of nucleolus-related inclusions. 消除核仁相关内含物的核蛋白质量控制系统。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2024-12-17 DOI: 10.1038/s44318-024-00333-9

Lorène Brunello, Jolanta Polanowska, Léo Le Tareau, Chantal Maghames, Virginie Georget, Charlotte Guette, Karima Chaoui, Stéphanie Balor, Marie-Françoise O'Donohue, Marie-Pierre Bousquet, Pierre-Emmanuel Gleizes, Dimitris P Xirodimas

{"title":"A nuclear protein quality control system for elimination of nucleolus-related inclusions.","authors":"Lorène Brunello, Jolanta Polanowska, Léo Le Tareau, Chantal Maghames, Virginie Georget, Charlotte Guette, Karima Chaoui, Stéphanie Balor, Marie-Françoise O'Donohue, Marie-Pierre Bousquet, Pierre-Emmanuel Gleizes, Dimitris P Xirodimas","doi":"10.1038/s44318-024-00333-9","DOIUrl":"10.1038/s44318-024-00333-9","url":null,"abstract":"The identification of pathways that control elimination of protein inclusions is essential to understand the cellular response to proteotoxicity, particularly in the nuclear compartment, for which our knowledge is limited. We report that stress-induced nuclear inclusions related to the nucleolus are eliminated upon stress alleviation during the recovery period. This process is independent of autophagy/lysosome and CRM1-mediated nuclear export pathways, but strictly depends on the ubiquitin-activating E1 enzyme, UBA1, and on nuclear proteasomes that are recruited into the formed inclusions. UBA1 activity is essential only for the recovery process but dispensable for nuclear inclusion formation. Furthermore, the E3 ligase HUWE1 and HSP70 are components of the ubiquitin/chaperone systems that promote inclusion elimination. The recovery process also requires RNA Pol I-dependent production of the lncRNA IGS42 during stress. IGS42 localises within the formed inclusions and promotes their elimination by preserving the mobility of resident proteins. These findings reveal a protein quality control system that operates within the nucleus for the elimination of stress-induced nucleolus-related inclusions.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"801-823"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11791210/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142848317","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Two FAM134B isoforms differentially regulate ER dynamics during myogenesis. 两种FAM134B异构体在肌发生过程中对内质网动力学的调节存在差异。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2025-01-06 DOI: 10.1038/s44318-024-00356-2

Viviana Buonomo, Kateryna Lohachova, Alessio Reggio, Sara Cano-Franco, Michele Cillo, Lucia Santorelli, Rossella Venditti, Elena Polishchuk, Ivana Peluso, Lorene Brunello, Carmine Cirillo, Sara Petrosino, Malan Silva, Rossella De Cegli, Sabrina Di Bartolomeo, Cesare Gargioli, Paolo Swuec, Mirko Cortese, Alexandra Stolz, Ramachandra M Bhaskara, Paolo Grumati

{"title":"Two FAM134B isoforms differentially regulate ER dynamics during myogenesis.","authors":"Viviana Buonomo, Kateryna Lohachova, Alessio Reggio, Sara Cano-Franco, Michele Cillo, Lucia Santorelli, Rossella Venditti, Elena Polishchuk, Ivana Peluso, Lorene Brunello, Carmine Cirillo, Sara Petrosino, Malan Silva, Rossella De Cegli, Sabrina Di Bartolomeo, Cesare Gargioli, Paolo Swuec, Mirko Cortese, Alexandra Stolz, Ramachandra M Bhaskara, Paolo Grumati","doi":"10.1038/s44318-024-00356-2","DOIUrl":"10.1038/s44318-024-00356-2","url":null,"abstract":"Endoplasmic reticulum (ER) plasticity and ER-phagy are intertwined processes essential for maintaining ER dynamics. We investigated the interplay between two isoforms of the ER-phagy receptor FAM134B in regulating ER remodeling in differentiating myoblasts. During myogenesis, the canonical FAM134B1 is degraded, while its isoform FAM134B2 is transcriptionally upregulated. The switch, favoring FAM134B2, is an important regulator of ER morphology during myogenesis. FAM134B2 partial reticulon homology domain, with its rigid conformational characteristics, enables efficient ER reshaping. FAM134B2 action increases in the active phase of differentiation leading to ER restructuring via ER-phagy, which then reverts to physiological levels when myotubes are mature and the ER is reorganized. Knocking out both FAM134B isoforms in myotubes results in an aberrant proteome landscape and the formation of dilated ER structures, both of which are rescued by FAM134B2 re-expression. Our results underscore how the fine-tuning of FAM134B isoforms and ER-phagy orchestrate the ER dynamics during myogenesis providing insights into the molecular mechanisms governing ER homeostasis in muscle cells.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":"44 4","pages":"1039-1073"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11832904/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143442568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

qTAG: an adaptable plasmid scaffold for CRISPR-based endogenous tagging. qTAG：用于基于crispr的内源性标记的适应性质粒支架。

IF 9.4 1区生物学

EMBO Journal Pub Date : 2025-02-01 Epub Date: 2024-12-12 DOI: 10.1038/s44318-024-00337-5

Reuben Philip, Amit Sharma, Laura Matellan, Anna C Erpf, Wen-Hsin Hsu, Johnny M Tkach, Haley D M Wyatt, Laurence Pelletier

{"title":"qTAG: an adaptable plasmid scaffold for CRISPR-based endogenous tagging.","authors":"Reuben Philip, Amit Sharma, Laura Matellan, Anna C Erpf, Wen-Hsin Hsu, Johnny M Tkach, Haley D M Wyatt, Laurence Pelletier","doi":"10.1038/s44318-024-00337-5","DOIUrl":"10.1038/s44318-024-00337-5","url":null,"abstract":"Endogenous tagging enables the study of proteins within their native regulatory context, typically using CRISPR to insert tag sequences directly into the gene sequence. Here, we introduce qTAG, a collection of repair cassettes that makes endogenous tagging more accessible. The cassettes support N- and C-terminal tagging with commonly used selectable markers and feature restriction sites for easy modification. Lox sites also enable the removal of the marker gene after successful integration. We demonstrate the utility of qTAG with a range of diverse tags for applications in fluorescence imaging, proximity labeling, epitope tagging, and targeted protein degradation. The system includes novel tags like mStayGold, offering enhanced brightness and photostability for live-cell imaging of native protein dynamics. Additionally, we explore alternative cassette designs for conditional expression tagging, selectable knockout tagging, and safe-harbor expression. The plasmid collection is available through Addgene, featuring ready-to-use constructs for common subcellular markers and tagging cassettes to target genes of interest. The qTAG system will serve as an open resource for researchers to adapt and tailor their own experiments.","PeriodicalId":50533,"journal":{"name":"EMBO Journal","volume":" ","pages":"947-974"},"PeriodicalIF":9.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11790981/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142819793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0