Max L B Grönloh, Werner J van der Meer, Merel E Tebbens, Jaap D van Buul
{"title":"Leukocyte transendothelial migration hotspots at a glance.","authors":"Max L B Grönloh, Werner J van der Meer, Merel E Tebbens, Jaap D van Buul","doi":"10.1242/jcs.263862","DOIUrl":"https://doi.org/10.1242/jcs.263862","url":null,"abstract":"<p><p>Leukocytes traverse the vasculature to reach sites of infection by sequentially crossing two distinct barriers - the endothelial barrier, during transendothelial migration (TEM), and the pericyte barrier, during trans-pericyte migration (TPM). Emerging evidence has underscored that TEM and TPM do not occur randomly but are confined to specialized 'hotspot' regions. This Cell Science at a Glance article and the accompanying poster overview the mechanisms underlying the heterogeneity within the inflamed endothelial monolayer, as well as within the perivascular cells, that defines these hotspots, highlighting how leukocytes themselves can actively induce new hotspots during the extravasation process. Furthermore, we explore the diverse mechanisms governing the production, storage and presentation of chemokines during extravasation. Finally, we elucidate the regulatory strategies employed by endothelial cells to maintain vascular integrity and prevent leakage during leukocyte transmigration, collectively offering a comprehensive perspective on the intricate interactions between leukocytes and (peri)vascular cells during immune cell extravasation.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":"138 11","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144317021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Correction: Defining the dynamin-based ring organizing center on the peroxisome-dividing machinery isolated from Cyanidioschyzon merolae.","authors":"Yuuta Imoto, Yuichi Abe, Kanji Okumoto, Masanori Honsho, Haruko Kuroiwa, Tsuneyoshi Kuroiwa, Yukio Fujiki","doi":"10.1242/jcs.264019","DOIUrl":"https://doi.org/10.1242/jcs.264019","url":null,"abstract":"","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":"138 10","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144093780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Cell deformations generated by stochastic actomyosin waves drive in vivo random-walk swimming migration.","authors":"Cyril Andrieu, Bren Hunyi Lee, Anna Franz","doi":"10.1242/jcs.263787","DOIUrl":"10.1242/jcs.263787","url":null,"abstract":"<p><p>Amoeboid cell migration drives many developmental and disease-related processes, including immune responses and cancer metastasis. Swimming migration is a subtype of amoeboid migration that is observed in cells in suspension ex vivo. However, the mechanism underlying swimming migration in vivo is unknown. Using Drosophila fat body cells (FBCs) as a model, we show that FBCs actively swim to patrol the pupa by random walk. Their migration is powered through actomyosin waves that exert compressive forces as they travel to the cell rear, causing cell deformations. Unlike in other types of amoeboid migration, Rho1 (the Drosophila orthologue of RhoA), Cdc42 and Rac1 are all required for regulation of formin-driven actin polymerization during FBC migration. We find that Rho1 at the cell rear induces actomyosin contractions via Rho kinase and myosin II. We show that contractile actin waves display a stochastic behaviour, inducing either cell elongation or rounding, suggesting that non-reciprocal cell deformations drive locomotion. Importantly, our work in a physiological system reveals that stochastic actomyosin waves promote random-walk swimming migration to enable fast, long-range cell dispersal. We propose that this individualist migration behaviour collectively allows patrolling of the pupal body.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148041/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kunlun Yin, Luke Villareal, Xiangxiang Wu, Mariella Arcos, Jordan Lee, David R Martin, Julie G In, Kimberly Leslie, Donna D Zhang, Xiang Xue
{"title":"The STEAP4 target NQO1 mediates colon tumorigenesis.","authors":"Kunlun Yin, Luke Villareal, Xiangxiang Wu, Mariella Arcos, Jordan Lee, David R Martin, Julie G In, Kimberly Leslie, Donna D Zhang, Xiang Xue","doi":"10.1242/jcs.263402","DOIUrl":"10.1242/jcs.263402","url":null,"abstract":"<p><p>Colorectal cancer (CRC) remains a major global health concern, necessitating advancements in therapeutic strategies. Understanding the mechanisms driving CRC is crucial for developing effective treatments. Previous studies, including our own, highlight the role of six-transmembrane epithelial antigen of prostate 4 (STEAP4) in promoting colon tumorigenesis through reactive oxygen species (ROS) generation, making it a promising target. Our research provides compelling evidence that STEAP4 knockout significantly reduces colon tumorigenesis in a genetically engineered mouse model. Suppressing STEAP4 via knockdown techniques effectively attenuated the nuclear factor erythroid 2-related factor 2 (NRF2)-NAD(P)H:quinone oxidoreductase 1 (NQO1) signaling pathway, inducing apoptosis and autophagy, leading to substantial reductions in xenograft tumor growth. In contrast, STEAP4 overexpression amplified ROS production and activated the NRF2-NQO1 pathway in a ferric iron (Fe3+)-dependent manner. Notably, bioactivatable drugs targeting NQO1 were highly effective at eradicating STEAP4-overexpressing colon cancer cells. These findings highlight the potential of targeted therapeutic interventions for CRC, particularly through STEAP4 modulation. In conclusion, our study advances understanding of the role of STEAP4 in colon tumorigenesis, offering promising avenues for novel CRC treatments.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148032/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144000981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Giulia Viola, Yasmeen W Ibrahim, Kyle A Jacobs, Joël Lemière, Matthew L Kutys, Torsten Wittmann
{"title":"Quantitative comparison of fluorescent proteins using protein nanocages in live cells.","authors":"Giulia Viola, Yasmeen W Ibrahim, Kyle A Jacobs, Joël Lemière, Matthew L Kutys, Torsten Wittmann","doi":"10.1242/jcs.263858","DOIUrl":"10.1242/jcs.263858","url":null,"abstract":"<p><p>To standardize comparison of fluorescent protein performance on a molecule-by-molecule basis in a physiological intracellular environment, we constructed fluorescent protein-tagged I3-01 peptides that self-assemble into stable 60-subunit dodecahedrons inside live mammalian cells. We were especially interested in determining which of the recently published monomeric StayGold variants is best for live microscopy in mammalian cells. Combining nanocage brightness and photobleaching measurements into a single metric, mStayGold stood out as far superior to all other green and red fluorescent proteins we tested with a functional lifetime that is at least 8-10-fold longer compared with EGFP or mEmerald. Analysis of intracellular nanocage diffusion further confirmed the monomeric nature of mStayGold, and we demonstrate that mStayGold-tagged nanocages can serve as highly photostable nanoparticles to analyze intracellular biophysical properties. Analysis of frequently used red fluorescent proteins was less encouraging and recent mScarlet or mRuby variants did not perform substantially better than mCherry on a typical spinning disc confocal microscope system, highlighting the importance of a standardized method to benchmark fluorescent proteins to make optimal choices for specific experimental setups.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148025/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Marc Severin, Rikke K Hansen, Michala G Rolver, Tove Hels, Kenji Maeda, Luis A Pardo, Stine F Pedersen
{"title":"Tumor acidosis supports cancer cell lipid uptake via a rapid transporter-independent mechanism.","authors":"Marc Severin, Rikke K Hansen, Michala G Rolver, Tove Hels, Kenji Maeda, Luis A Pardo, Stine F Pedersen","doi":"10.1242/jcs.263688","DOIUrl":"10.1242/jcs.263688","url":null,"abstract":"<p><p>Tumor acidosis alters cancer cell metabolism and favors aggressive disease progression. Cancer cells in acidic environments increase lipid droplet accumulation and oxidative phosphorylation, which are characteristics of aggressive cancers. Here, we used live imaging, shotgun lipidomics and immunofluorescence analyses of mammary and pancreatic cancer cells to demonstrate that both acute acidosis and adaptation to acidic growth drive rapid uptake of fatty acids (FAs), which are converted to triacylglycerols and stored in lipid droplets. Consistent with being independent of de novo synthesis, triacylglycerol and lipid droplet accumulation in acid-adapted cells was unaffected by FA synthetase (FAS, encoded by FASN) inhibitors. Macropinocytosis, which is upregulated in acid-adapted cells, partially contributed to FA uptake, which was independent of other protein-facilitated lipid uptake mechanisms, including uptake via CD36 and FATP2, and caveolin- and clathrin-dependent endocytosis. We propose that a major mechanism by which tumor acidosis drives FA uptake is through neutralizing protonation of negatively charged FAs allowing their diffusive, transporter-independent uptake. We suggest that this could be a major factor triggering acidosis-driven metabolic rewiring.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143795577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hyunjeong Yoo, Taehyun Kim, Sungjin Ryu, Donghee Ko, Jeesoo Kim, Hee-Jung Choi, Yongdae Shin, Kunsoo Rhee
{"title":"Enhancement of CEP215 dynamics for spindle pole assembly during mitosis.","authors":"Hyunjeong Yoo, Taehyun Kim, Sungjin Ryu, Donghee Ko, Jeesoo Kim, Hee-Jung Choi, Yongdae Shin, Kunsoo Rhee","doi":"10.1242/jcs.263542","DOIUrl":"10.1242/jcs.263542","url":null,"abstract":"<p><p>The microtubule-organizing activity of centrosomes fluctuates during the cell cycle, reaching the highest levels at M phase. CEP215 (also known as CDK5RAP2) is a key pericentriolar material (PCM) protein for microtubule organization of the human centrosome. Here, we provide evidence that CEP215 exhibits a dynamically suppressed, solid-like state in interphase centrosomes, and becomes a more dynamic state in mitotic centrosomes. Specific interaction with PCNT, another centrosome protein, is crucial for diffusible molecular dynamicity of the CEP215 protein. We also found that the cluster formation activity of CEP215 is impaired in a light-inducible system when its coiled-coil domains (CCDs) are truncated. Defects in spindle pole assembly and spindle formation were accompanied in the cells whose CEP215 is replaced with the CCD-truncated mutants. Our results support the notion that the diffusible mobility of CEP215 is enhanced by both homotypic and heterotypic interactions among CCDs, especially at mitotic spindle poles. This work highlights that biophysical properties of the PCM proteins at the centrosomes fluctuate during the cell cycle.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148038/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ting-Wei Chang, Ming-Chieh Lin, Chia-Jung Yu, Fang-Jen S Lee
{"title":"The phosphorylation of Pak1 by Erk1/2 to drive cell migration requires Arl4D acting as a scaffolding protein.","authors":"Ting-Wei Chang, Ming-Chieh Lin, Chia-Jung Yu, Fang-Jen S Lee","doi":"10.1242/jcs.263812","DOIUrl":"10.1242/jcs.263812","url":null,"abstract":"<p><p>Activation of extracellular signal-regulated kinases 1 and 2 (Erk1/2; also known as MAPK3 and MAPK1, respectively) at the plasma membrane usually leads to their translocation to various intracellular sites, where scaffolding proteins mediate substrate targeting. However, in platelet-derived growth factor (PDGF)-induced signaling, Erk1/2 phosphorylate Pak1 to drive cell migration while remaining at the plasma membrane, raising the question of whether scaffolding proteins are required. Similarly, the small GTPase Arf-like protein 4D (Arl4D) promotes cell migration by recruiting Pak1 to the plasma membrane and facilitating its phosphorylation, although the mechanism linking recruitment to phosphorylation remains unclear. To address these questions, we show that Arl4D functions as a scaffolding protein by recruiting Erk1/2 and Pak1 to the plasma membrane, assembling them into a functional complex. This complex allows Erk1/2 to phosphorylate Pak1, supporting the role of the latter in cell migration. Our findings identify Arl4D as a novel regulator of Erk1/2, reveal a conserved role of scaffolding proteins in Erk1/2 substrate targeting, and uncover an unrecognized interplay among Arl4D, Erk1/2 and Pak1. These insights provide a deeper understanding of the molecular coordination underlying Pak1-mediated cell migration and its regulation by Erk1/2 and Arl4D.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148019/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002408","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"PI31 is a positive regulator of 20S immunoproteasome assembly.","authors":"Jason Wang, Abbey Kjellgren, George N DeMartino","doi":"10.1242/jcs.263887","DOIUrl":"10.1242/jcs.263887","url":null,"abstract":"<p><p>Proteasome inhibitor of 31,000 Da (PI31) is a 20S proteasome-binding protein originally identified as an inhibitor of in vitro 20S proteasome activity. Although recent studies have elucidated a detailed structural basis for this inhibitory activity, the physiological significance of PI31-mediated proteasome inhibition remains uncertain, and multiple alternative cellular roles for PI31 have been described. Here, we report a role for PI31 as a positive regulator for assembly of the 20S immunoproteasome (20Si), a compositionally and functionally distinct isoform of the proteasome that is poorly inhibited by PI31. Genetic ablation of PI31 in mammalian cells had no effect on the cellular content or activity of constitutively expressed proteasomes but reduced the cellular content and activity of interferon-γ-induced immunoproteasomes. This selective effect results from impaired 20Si assembly, evidenced by the accumulation of 20Si assembly intermediates. Our results highlight a distinction in the assembly pathways of constitutive proteasomes and immunoproteasomes and indicate that PI31 plays a chaperone-like role in the selective assembly of 20S immunoproteasomes.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148024/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kirti Prakash, David Baddeley, Christian Eggeling, Reto Fiolka, Rainer Heintzmann, Suliana Manley, Aleksandra Radenovic, Hari Shroff, Carlas Smith, Lothar Schermelleh
{"title":"Resolution in super-resolution microscopy - facts, artifacts, technological advancements and biological applications.","authors":"Kirti Prakash, David Baddeley, Christian Eggeling, Reto Fiolka, Rainer Heintzmann, Suliana Manley, Aleksandra Radenovic, Hari Shroff, Carlas Smith, Lothar Schermelleh","doi":"10.1242/jcs.263567","DOIUrl":"10.1242/jcs.263567","url":null,"abstract":"<p><p>Super-resolution microscopy (SRM) has undeniable potential for scientific discovery, yet still presents many challenges that hinder its widespread adoption, including technical trade-offs between resolution, speed and photodamage, as well as limitations in imaging live samples and larger, more complex biological structures. Furthermore, SRM often requires specialized expertise and complex instrumentation, which can deter biologists from fully embracing the technology. In this Perspective, a follow-up to our recent Q&A article, we aim to demystify these challenges by addressing common questions and misconceptions surrounding SRM. Experts offer practical insights into how biologists can maximize the benefits of SRM while navigating issues such as photobleaching, image artifacts and the limitations of existing techniques. We also highlight recent developments in SRM that continue to push the boundaries of resolution. Our goal is to equip researchers with the crucial knowledge they need to harness the full potential of SRM.</p>","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":"138 10","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12148039/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144150416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}