Journal of cell science最新文献_第4页

PIKFYVE inhibition induces endosome- and lysosome-derived vacuole enlargement via ammonium accumulation. 抑制 PIKFYVE 可通过铵积累诱导内膜体和溶酶体衍生的液泡增大。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-26 DOI: 10.1242/jcs.262236

Junsuke Uwada, Hitomi Nakazawa, Takeshi Kiyoi, Takashi Yazawa, Ikunobu Muramatsu, Takayoshi Masuoka

{"title":"PIKFYVE inhibition induces endosome- and lysosome-derived vacuole enlargement via ammonium accumulation.","authors":"Junsuke Uwada, Hitomi Nakazawa, Takeshi Kiyoi, Takashi Yazawa, Ikunobu Muramatsu, Takayoshi Masuoka","doi":"10.1242/jcs.262236","DOIUrl":"https://doi.org/10.1242/jcs.262236","url":null,"abstract":"FYVE-type zinc finger-containing phosphoinositide kinase (PIKFYVE), that is essential for PtdIns(3,5)P2 production, is an important regulator of lysosomal homeostasis. PIKFYVE dysfunction leads to cytoplasmic vacuolization; however, the underlying mechanism remains unknown. In this study, we explored the cause of vacuole enlargement upon PIKFYVE inhibition in DU145 prostate cancer cells. Enlargement of vacuoles by PIKFYVE inhibition required glutamine and its metabolism by glutaminases. Addition of ammonia, a metabolite of glutamine, was sufficient to enlarge vacuoles via PIKFYVE inhibition. Moreover, PIKFYVE inhibition led to intracellular ammonium accumulation. Endosome-lysosome permeabilization resulted in ammonium leakage from the cells, indicating ammonium accumulation in the endosomes and lysosomes. Ammonium accumulation and vacuole expansion were suppressed by the lysosomal lumen neutralization. It is therefore assumed that PIKFYVE inhibition interferes with the efflux of NH4+, which is protonated NH3 in the lysosomal lumen, leading to osmotic swelling of vacuoles. Notably, glutamine or ammonium is required for PIKFYVE inhibition-induced suppression of lysosomal function and autophagic flux. In conclusion, this study showed that PIKfyve inhibition disrupts lysosomal homeostasis via ammonium accumulation.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142716300","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}

引用次数: 0

Phosphorylation on serine 72 modulates Rab7A palmitoylation and retromer recruitment. 丝氨酸 72 上的磷酸化调节 Rab7A 棕榈酰化和 retromer 招募。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-25 DOI: 10.1242/jcs.262177

Graziana Modica, Laura Tejeda-Valencia, Etienne Sauvageau, Seda Yasa, Juliette Maes, Olga Skorobogata, Stephane Lefrancois

{"title":"Phosphorylation on serine 72 modulates Rab7A palmitoylation and retromer recruitment.","authors":"Graziana Modica, Laura Tejeda-Valencia, Etienne Sauvageau, Seda Yasa, Juliette Maes, Olga Skorobogata, Stephane Lefrancois","doi":"10.1242/jcs.262177","DOIUrl":"https://doi.org/10.1242/jcs.262177","url":null,"abstract":"Rab7A has a key role in regulating membrane trafficking at late endosomes. By interacting with several different effectors, this small GTPase controls late endosome mobility, orchestrates fusion events between late endosomes and lysosomes, and participates in the formation of and regulates the fusion between autophagosomes and lysosomes. Rab7A is also responsible for the spatiotemporal recruitment of retromer, which is required for the endosome-to-TGN retrieval of cargo-receptors such as sortilin and CI-MPR. Recently several post-translational modifications have been shown to modulate Rab7A functions, including palmitoylation, ubiquitination and phosphorylation. Here we show that phosphorylation of Rab7A at serine 72 is important to modulate its interaction with retromer, as the non-phosphorylatable Rab7AS72A mutant is not able to interact with and recruit retromer to late endosomes. We have previously shown that Rab7A palmitoylation is also required for efficient retromer recruitment. We found that palmitoylation of Rab7AS72A is reduced compared to the wild-type protein, suggesting an interplay between S72 phosphorylation and palmitoylation in regulating the Rab7A/retromer interaction. Finally, we identify NEK7 as a kinase required to phosphorylate Rab7A to promote retromer binding and recruitment.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142709578","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}

引用次数: 0

AMPK associates with and causes fragmentation of the Golgi by phosphorylating the guanine nucleotide exchange factor GBF1. AMPK 通过磷酸化鸟嘌呤核苷酸交换因子 GBF1 与高尔基体结合并导致其破碎。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-22 DOI: 10.1242/jcs.262182

Jordana B Freemantle, Mhairi C Towler, Emma R Hudson, Thomas Macartney, Monika Zwirek, David J K Liu, David A Pan, Sreenivasan Ponnambalam, D Grahame Hardie

{"title":"AMPK associates with and causes fragmentation of the Golgi by phosphorylating the guanine nucleotide exchange factor GBF1.","authors":"Jordana B Freemantle, Mhairi C Towler, Emma R Hudson, Thomas Macartney, Monika Zwirek, David J K Liu, David A Pan, Sreenivasan Ponnambalam, D Grahame Hardie","doi":"10.1242/jcs.262182","DOIUrl":"https://doi.org/10.1242/jcs.262182","url":null,"abstract":"AMPK is an energy sensor that regulates cellular functions in response to changes in energy availability. However, whether AMPK activity is spatially regulated, and the implications for cell function, have been unclear. We now report that AMPK associates with the Golgi apparatus, and that its activation by two specific pharmacological activators leads to Golgi fragmentation similar to that caused by the antibiotic Golgicide A, an inhibitor of Golgi-specific Brefeldin A resistance Factor-1 (GBF1), a guanine nucleotide exchange factor that targets ADP-Ribosylation Factor-1 (ARF1). Golgi fragmentation in response to AMPK activators is lost in cells carrying gene knockouts of AMPK-a subunits. AMPK has been previously reported to phosphorylate GBF1 at residue Thr1337, and its activation causes phosphorylation at that residue. Importantly, Golgi disassembly upon AMPK activation is blocked in cells expressing a non-phosphorylatable GBF1-T1337A mutant generated by gene editing. Furthermore, the trafficking of a plasma membrane-targetted protein through the Golgi complex is delayed by AMPK activation. Our findings provide a mechanism to link AMPK activation during cellular energy stress to down-regulation of protein trafficking involving the Golgi apparatus.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142687210","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}

引用次数: 0

Chemomechanical regulation of EZH2 localization controls epithelial-mesenchymal transition. EZH2定位的化学机械调控控制着上皮-间质转化。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-25 DOI: 10.1242/jcs.262190

Jessica L Sacco, Zachary T Vaneman, Ava Self, Elix Sumner, Stella Kibinda, Chinmay S Sankhe, Esther W Gomez

{"title":"Chemomechanical regulation of EZH2 localization controls epithelial-mesenchymal transition.","authors":"Jessica L Sacco, Zachary T Vaneman, Ava Self, Elix Sumner, Stella Kibinda, Chinmay S Sankhe, Esther W Gomez","doi":"10.1242/jcs.262190","DOIUrl":"10.1242/jcs.262190","url":null,"abstract":"The methyltransferase enhancer of zeste homolog 2 (EZH2) regulates gene expression, and aberrant EZH2 expression and signaling can drive fibrosis and cancer. However, it is not clear how chemical and mechanical signals are integrated to regulate EZH2 and gene expression. We show that culture of cells on stiff matrices in concert with transforming growth factor (TGF)-β1 promotes nuclear localization of EZH2 and an increase in the levels of the corresponding histone modification, H3K27me3, thereby regulating gene expression. EZH2 activity and expression are required for TGFβ1- and stiffness-induced increases in H3K27me3 levels as well as for morphological and gene expression changes associated with epithelial-mesenchymal transition (EMT). Inhibition of Rho associated kinase (ROCK) proteins or myosin II signaling attenuates TGFβ1-induced nuclear localization of EZH2 and decreases H3K27me3 levels in cells cultured on stiff substrata, suggesting that cellular contractility, in concert with a major cancer signaling regulator TGFβ1, modulates EZH2 subcellular localization. These findings provide a contractility-dependent mechanism by which matrix stiffness and TGFβ1 together mediate EZH2 signaling to promote EMT.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142501120","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}

引用次数: 0

ImmunoCellCycle-ID - a high-precision immunofluorescence-based method for cell cycle identification. ImmunoCellCycle-ID--一种基于免疫荧光的高精度细胞周期鉴定方法。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-20 DOI: 10.1242/jcs.263414

Yu-Lin Chen, Yu-Chia Chen, Aussie Suzuki

引用次数: 0

DRAK2 regulates myosin light chain phosphorylation in T cells. DRAK2 可调节 T 细胞中肌球蛋白轻链的磷酸化。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-20 DOI: 10.1242/jcs.261813

Benjamin A Wilander, Tarsha L Harris, Alexandra H Mandarano, Cliff S Guy, Mollie S Prater, Shondra M Pruett-Miller, Stacey K Ogden, Maureen A McGargill

{"title":"DRAK2 regulates myosin light chain phosphorylation in T cells.","authors":"Benjamin A Wilander, Tarsha L Harris, Alexandra H Mandarano, Cliff S Guy, Mollie S Prater, Shondra M Pruett-Miller, Stacey K Ogden, Maureen A McGargill","doi":"10.1242/jcs.261813","DOIUrl":"10.1242/jcs.261813","url":null,"abstract":"Death-associated protein kinase-related apoptosis-inducing kinase-2 (DRAK2; also known as STK17B) is a serine/threonine kinase expressed in T cells. Drak2-deficient (Drak2-/-) mice respond effectively to tumors and pathogens while displaying resistance to T cell-mediated autoimmune disease. However, the molecular mechanisms by which DRAK2 impacts T cell function remain unclear. Gaining further insight into the function of DRAK2 in T cells will shed light on differentially regulated pathways in autoreactive and pathogen-specific T cells, which is crucial for improving autoimmune therapies. Here, we demonstrate that DRAK2 contributes to activation of myosin light chain (MLC2, encoded by Myl2) in both murine and human T cells. In the absence of Drak2, the amount of polymerized actin was decreased, suggesting that DRAK2 modulates actomyosin dynamics. We further show that myosin-dependent T cell functions, such as migration, T cell receptor microcluster accumulation, and conjugation to antigen presenting cells are decreased in the absence of Drak2. These findings reveal that DRAK2 plays an important role in regulating MLC activation within T cells.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11607690/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466387","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}

引用次数: 0

The lysosomal lipid transporter LIMP-2 is part of lysosome-ER STARD3-VAPB-dependent contact sites. 溶酶体脂质转运体 LIMP-2/SCARB2 是溶酶体-内质网 STARD3-VAPB 依赖性接触点的一部分。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-26 DOI: 10.1242/jcs.261810

Sönke Rudnik, Saskia Heybrock, Etienne Coyaud, Zizhen Xu, Dante Neculai, Brian Raught, Viola Oorschot, Cecilia Heus, Judith Klumperman, Paul Saftig

{"title":"The lysosomal lipid transporter LIMP-2 is part of lysosome-ER STARD3-VAPB-dependent contact sites.","authors":"Sönke Rudnik, Saskia Heybrock, Etienne Coyaud, Zizhen Xu, Dante Neculai, Brian Raught, Viola Oorschot, Cecilia Heus, Judith Klumperman, Paul Saftig","doi":"10.1242/jcs.261810","DOIUrl":"10.1242/jcs.261810","url":null,"abstract":"LIMP-2 (also known as SCARB2) is an abundant lysosomal membrane protein. Previous studies have shown that LIMP-2 functions as a virus receptor, a chaperone for lysosomal enzyme targeting and a lipid transporter. The large luminal domain of LIMP-2 contains a hydrophobic tunnel that enables transport of phospholipids, sphingosine and cholesterol from the lysosomal lumen to the membrane. The question about the fate of the lipids after LIMP-2-mediated transport is largely unexplored. To elucidate whether LIMP-2 is present at contact sites between lysosomes and the endoplasmic reticulum (ER), we performed a proximity-based interaction screen. This revealed that LIMP-2 interacts with the endosomal protein STARD3 and the ER-resident protein VAPB. Using imaging and co-immunoprecipitation, we demonstrated colocalization and physical interaction between LIMP-2 and these proteins. Moreover, we found that interaction of LIMP-2 with VAPB required the presence of STARD3. Our findings suggest that LIMP-2 is present at ER-lysosome contact sites, possibly facilitating cholesterol transport from the lysosomal to the ER membrane. This suggests a novel mechanism for inter-organelle communication and lipid trafficking mediated by LIMP-2.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142380944","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}

引用次数: 0

HIV-1 N-myristoylation-dependent hijacking of late endosomes/lysosomes to drive Gag assembly in macrophages. HIV-1 N-肉豆蔻酰化依赖性劫持晚期内体/溶酶体，以驱动巨噬细胞中 Gag 的组装。

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-21 DOI: 10.1242/jcs.263588

Gabriel I Guajardo-Contreras, Ana L Abdalla, Alex Chen, Meijuan Niu, Erwan Beauchamp, Luc G Berthiaume, Alan W Cochrane, Andrew J Mouland

{"title":"HIV-1 N-myristoylation-dependent hijacking of late endosomes/lysosomes to drive Gag assembly in macrophages.","authors":"Gabriel I Guajardo-Contreras, Ana L Abdalla, Alex Chen, Meijuan Niu, Erwan Beauchamp, Luc G Berthiaume, Alan W Cochrane, Andrew J Mouland","doi":"10.1242/jcs.263588","DOIUrl":"10.1242/jcs.263588","url":null,"abstract":"Macrophages represent an important viral reservoir in HIV-1-infected individuals. Different from T cells, HIV-1 assembly in macrophages occurs at intracellular compartments termed virus-containing compartments (VCCs). Our previous research in HeLa cells - in which assembly resembles that found in infected T cells - suggested that late endosomes/lysosomes (LELs) play a role in HIV-1 trafficking towards its assembly sites. However, the role of LELs during assembly at VCCs is not fully understood. Herein, we used the HIV-1-inducible cell line THP-1 GagZip as a model to study HIV-1 Gag intracellular trafficking and assembly in macrophages. We demonstrated LEL involvement at VCCs using various microscopy techniques and biochemical approaches. Live-cell imaging revealed that HIV-1 repositions LELs towards the plasma membrane and modulates their motility. We showed that Arl8b-mediated LEL repositioning is not responsible for Gag trafficking to VCCs. Additionally, the inhibition of myristoylation by PCLX-001 decreased the presence of Gag on endosomes and inhibited VCC formation in both the THP-1 cell line and primary macrophages. In conclusion, we present evidence supporting the idea that HIV-1 manipulates the LEL trajectory to guide Gag to VCCs in an N-myristoylation-dependent manner.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11607699/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142501121","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}

引用次数: 0

CENP-C-targeted PLK-1 regulates kinetochore function in C. elegans embryos. CENP-C靶向PLK-1调控优雅子胚胎中的动点功能

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-28 DOI: 10.1242/jcs.262327

Laura Bel Borja, Samuel J P Taylor, Flavie Soubigou, Federico Pelisch

{"title":"CENP-C-targeted PLK-1 regulates kinetochore function in C. elegans embryos.","authors":"Laura Bel Borja, Samuel J P Taylor, Flavie Soubigou, Federico Pelisch","doi":"10.1242/jcs.262327","DOIUrl":"10.1242/jcs.262327","url":null,"abstract":"Polo-like kinase 1 (PLK-1) is present in centrosomes, the nuclear envelope and kinetochores and plays a significant role in meiosis and mitosis. PLK-1 depletion or inhibition has severe consequences for spindle assembly, spindle assembly checkpoint (SAC) activation, chromosome segregation and cytokinesis. BUB-1 targets PLK-1 to the outer kinetochore and, in mammals, the inner kinetochore PLK1 targeting is mediated by the constitutive centromere associated network (CCAN). BUB-1-targeted PLK-1 plays a key role in SAC activation and has a SAC-independent role through targeting CDC-20. In contrast, whether there is a specific, non-redundant role for inner kinetochore targeted PLK-1 is unknown. Here, we used the Caenorhabditis elegans embryo to study the role of inner kinetochore PLK-1. We found that CENP-C, the sole CCAN component in C. elegans and other species, targets PLK-1 to the inner kinetochore during prometaphase and metaphase. Disruption of the CENP-C-PLK-1 interaction leads to an imbalance in kinetochore components and a defect in chromosome congression, without affecting CDC-20 recruitment. These findings indicate that PLK-1 kinetochore recruitment by CENP-C has at least partially distinct functions from outer kinetochore PLK-1, providing a platform for a better understanding of the different roles played by PLK-1 during mitosis.","PeriodicalId":15227,"journal":{"name":"Journal of cell science","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11634037/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142361604","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}

引用次数: 0

Dynamic remodelling of the endoplasmic reticulum for mitosis. 有丝分裂过程中内质网的动态重塑

IF 3.3 3区生物学

Journal of cell science Pub Date : 2024-11-15 Epub Date: 2024-11-25 DOI: 10.1242/jcs.261444

Suzan Kors, Anne-Lore Schlaitz

引用次数: 0