The Journal of Cell Biology最新文献_第8页

Recycling of cell surface membrane proteins from yeast endosomes is regulated by ubiquitinated Ist1 酵母内体的细胞膜蛋白的再循环是由泛素化的Ist1调控的

The Journal of Cell Biology Pub Date : 2021-06-19 DOI: 10.1101/2021.06.19.449106

K. Laidlaw, G. Calder, C. MacDonald

{"title":"Recycling of cell surface membrane proteins from yeast endosomes is regulated by ubiquitinated Ist1","authors":"K. Laidlaw, G. Calder, C. MacDonald","doi":"10.1101/2021.06.19.449106","DOIUrl":"https://doi.org/10.1101/2021.06.19.449106","url":null,"abstract":"Trafficking of cell surface membrane proteins to and from the plasma membrane impinges on myriad biological processes and ensures correct cellular function. Upon internalization, many surface proteins are recycled back to the plasma membrane. Although these endosomal trafficking pathways control surface protein activity, the precise regulatory features and division of labour between interconnected pathways is poorly defined. Furthermore, how well endosomal trafficking mechanisms are conserved is unclear. In yeast, we show cargo recycling back to the surface occurs through distinct pathways. In addition to retrograde recycling pathways via the late Golgi, used by synaptobrevins and driven by cargo ubiquitination, we find nutrient transporter recycling bypasses the Golgi in a pathway driven by cargo deubiquitination. Nutrient transporters rapidly internalize to, and recycle from, endosomes marked by Vps4 and the ESCRT-III associated factor Ist1. This compartment serves as both ‘early’ and ‘recycling’ endosome, implying these features are evolutionarily conserved. Ist1 has previously been implicated in recycling in yeast and other eukaryotes. We show Ist1 is ubiquitinated and this is required for proper endosomal recruitment and cargo recycling to the surface. Additionally, the ubiquitin-binding adaptor Npl4 and the essential ATPase Cdc48 are required for cargo recycling possibly through regulation of ubiquitinated Ist1. This collectively suggests mechanistic features of recycling from endosomes to the plasma membrane are also conserved.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122083748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Interphase microtubule disassembly is a signaling cue that drives cell rounding at mitotic entry 间期微管解体是一个信号提示，驱动细胞在有丝分裂进入时成圆

The Journal of Cell Biology Pub Date : 2021-06-14 DOI: 10.1083/jcb.202109065

Kévin Leguay, Barbara Decelle, Islam E. Elkholi, M. Bouvier, J. Côté, Sébastien Carréno

{"title":"Interphase microtubule disassembly is a signaling cue that drives cell rounding at mitotic entry","authors":"Kévin Leguay, Barbara Decelle, Islam E. Elkholi, M. Bouvier, J. Côté, Sébastien Carréno","doi":"10.1083/jcb.202109065","DOIUrl":"https://doi.org/10.1083/jcb.202109065","url":null,"abstract":"Reorganization of the cortical actin cytoskeleton at mitotic entry is essential to increase membrane tension for cell rounding1,2. This spherical shape is necessary for the biogenesis and organization of the mitotic spindle2-6. Proteins of the Ezrin, Radixin, Moesin (ERM) family play essential roles in mitotic morphogenesis by linking actomyosin forces to the plasma membrane2,3,7-10. While ERMs drive metaphase cell rounding, the cell-cycle signals that prompt their conformational activation in mitosis are unknown11. We screened a library of small molecules using novel ERM biosensors12 and we unexpectedly found that drugs that disassemble microtubules promote ERM activation. Remarkably, cells disassemble their interphase microtubules while entering mitosis13. We further discovered that this disassembly of microtubules acts as a cell-cycle signal that directs ERM activation and metaphase cell rounding. We show that GEF-H1, a Rho-GEF inhibited by microtubule binding, acts downstream of microtubule disassembly to activate ERMs via RhoA and its kinase effector SLK. In addition, we demonstrate that GEF-H1 and Ect2, another Rho-GEF responsible for the generation of mitotic actomyosin forces6,14, act together to drive metaphase ERM activation and cell rounding. In summary, we report microtubule disassembly as a cell cycle signal that triggers a signaling network ensuring that actomyosin forces are efficiently integrated at the plasma membrane to promote cell rounding at mitotic entry.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"221 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131315110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

ER-lysosome lipid transfer protein VPS13C/PARK23 prevents aberrant mtDNA-dependent STING signaling er溶酶体脂质转移蛋白VPS13C/PARK23阻止异常mtdna依赖性STING信号传导

The Journal of Cell Biology Pub Date : 2021-06-08 DOI: 10.1101/2021.06.08.447593

William Hancock-Cerutti, Zheng Wu, A. Tharkeshwar, S. Ferguson, G. Shadel, P. De Camilli

引用次数: 26

WASP integrates substrate topology and cell polarity to guide neutrophil migration WASP整合了基质拓扑结构和细胞极性来引导中性粒细胞迁移

The Journal of Cell Biology Pub Date : 2021-05-14 DOI: 10.1101/2021.05.12.443722

Rachel M. Brunetti, Gabriele Kockelkoren, Preethi Raghavan, George R. R. Bell, Derek Britain, Natasha Puri, Sean R. Collins, M. Leonetti, D. Stamou, O. Weiner

{"title":"WASP integrates substrate topology and cell polarity to guide neutrophil migration","authors":"Rachel M. Brunetti, Gabriele Kockelkoren, Preethi Raghavan, George R. R. Bell, Derek Britain, Natasha Puri, Sean R. Collins, M. Leonetti, D. Stamou, O. Weiner","doi":"10.1101/2021.05.12.443722","DOIUrl":"https://doi.org/10.1101/2021.05.12.443722","url":null,"abstract":"To control their shape and movement, cells leverage nucleation promoting factors (NPFs) to regulate when and where they polymerize actin. Here we investigate the role of the immune-specific NPF WASP during neutrophil migration. Endogenously-tagged WASP localizes to substrate-induced plasma membrane deformations. Super-resolution imaging of live cells reveals that WASP preferentially enriches to the necks of these substrate-induced membrane invaginations, a distribution that could support substrate pinching. Unlike other curvature-sensitive proteins, WASP only enriches to membrane deformations at the cell front, where it controls Arp2/3 complex recruitment and actin polymerization. Despite relatively normal migration on flat substrates, WASP depletion causes defects in topology sensing and directed migration on textured substrates. WASP therefore both responds to and reinforces cell polarity during migration. Surprisingly, front-biased WASP puncta continue to form in the absence of Cdc42. We propose that WASP integrates substrate topology with cell polarity for 3D guidance by selectively polymerizing actin around substrate-induced membrane deformations at the leading edge. A misregulation of WASP-mediated contact guidance could provide insight into the immune disorder Wiskott-Aldrich syndrome.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"140 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114406090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 20

CRISPR screens for lipid regulators reveal a role for ER-bound SNX13 in lysosomal cholesterol export CRISPR筛选脂质调节因子揭示er结合SNX13在溶酶体胆固醇输出中的作用

The Journal of Cell Biology Pub Date : 2021-05-10 DOI: 10.1101/2021.05.10.443492

Albert Lu, F. Hsieh, C. Enrich, S. Pfeffer

{"title":"CRISPR screens for lipid regulators reveal a role for ER-bound SNX13 in lysosomal cholesterol export","authors":"Albert Lu, F. Hsieh, C. Enrich, S. Pfeffer","doi":"10.1101/2021.05.10.443492","DOIUrl":"https://doi.org/10.1101/2021.05.10.443492","url":null,"abstract":"We report here two genome-wide CRISPR screens carried out to identify genes that when knocked out, alter levels of lysosomal cholesterol or bis(monoacylglycero)phosphate. In addition, these screens were also carried out under conditions of NPC1 inhibition to identify modifiers of NPC1 function in lysosomal cholesterol export. The screens confirm tight co- regulation of cholesterol and bis(monoacylglycero)phosphate levels in cells, and reveal an unexpected role for the ER-localized, SNX13 protein as a negative regulator of lysosomal cholesterol export. In the absence of NPC1 function, SNX13 knockout decreases lysosomal cholesterol, and is accompanied by triacylglycerol-rich lipid droplet accumulation and increased lysosomal bis(monoacylglycero)phosphate. These experiments provide unexpected insight into the regulation of lysosomal lipids and modification of these processes by novel gene products. SUMMARY Genome-wide CRISPR screens carried out with and without NPC1 function identify shared pathways that coordinately control lysosomal cholesterol and bis(monoacylglycero)phosphate. ER-localized SNX13 protein plays an unexpected regulatory role in modifying NPC1 function to regulate cellular cholesterol localization.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"413 9","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134290001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

Atg39 links and deforms the outer and inner nuclear membranes in selective autophagy of the nucleus 在核的选择性自噬中，Atg39连接并变形核膜的内外膜

The Journal of Cell Biology Pub Date : 2021-03-29 DOI: 10.1101/2021.03.29.437603

Keisuke Mochida, Toshifumi Otani, Yuto Katsumata, Hiromi Kirisako, Chika Kakuta, T. Kotani, H. Nakatogawa

{"title":"Atg39 links and deforms the outer and inner nuclear membranes in selective autophagy of the nucleus","authors":"Keisuke Mochida, Toshifumi Otani, Yuto Katsumata, Hiromi Kirisako, Chika Kakuta, T. Kotani, H. Nakatogawa","doi":"10.1101/2021.03.29.437603","DOIUrl":"https://doi.org/10.1101/2021.03.29.437603","url":null,"abstract":"In selective autophagy of the nucleus (hereafter nucleophagy), nucleus-derived double membrane vesicles (NDVs) are formed, sequestered within autophagosomes, and delivered to lysosomes or vacuoles for degradation. In Saccharomyces cerevisiae, the nuclear envelope (NE) protein Atg39 acts as a nucleophagy receptor, which interacts with Atg8 to target NDVs to forming autophagosomal membranes. In this study, we revealed that Atg39 is anchored to the outer nuclear membrane (ONM) via its transmembrane domain and also associated with the inner nuclear membrane (INM) via membrane-binding amphipathic helices (APHs) in its perinuclear space region, thereby linking these membranes. We also revealed that overaccumulation of Atg39 causes the NE to protrude towards the cytoplasm, and the tips of the protrusions are pinched off to generate NDVs. The APHs of Atg39 are crucial for Atg39 assembly in the NE and subsequent NE protrusion. These findings suggest that the nucleophagy receptor Atg39 plays pivotal roles in NE deformation during the generation of NDVs to be degraded by nucleophagy.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"125 3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124269387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Spatiotemporal control of actomyosin contractility by MRCKβ signaling drives phagocytosis MRCKβ信号传导对肌动球蛋白收缩性的时空调控驱动吞噬作用

The Journal of Cell Biology Pub Date : 2021-03-25 DOI: 10.1101/2021.03.25.436833

C. Zihni, A. Georgiadis, Conor M. Ramsden, Elena Sánchez-Heras, B. Nommiste, O. Semenyuk, J. Bainbridge, P. Coffey, Alexander J. Smith, R. Ali, M. Balda, K. Matter

{"title":"Spatiotemporal control of actomyosin contractility by MRCKβ signaling drives phagocytosis","authors":"C. Zihni, A. Georgiadis, Conor M. Ramsden, Elena Sánchez-Heras, B. Nommiste, O. Semenyuk, J. Bainbridge, P. Coffey, Alexander J. Smith, R. Ali, M. Balda, K. Matter","doi":"10.1101/2021.03.25.436833","DOIUrl":"https://doi.org/10.1101/2021.03.25.436833","url":null,"abstract":"Phagocytosis requires myosin-generated contractile force to regulate actin dynamics. However, little is known about the molecular mechanisms that guide this complex morphodynamic process. Here we show that particle binding to Mer Tyrosine Kinase (MerTK), a widely expressed phagocytic receptor, stimulates phosphorylation of the Cdc42 GEF Dbl3 in the retinal pigment epithelium (RPE), triggering activation of MRCKβ and its co-effector N-WASP that cooperate to deform the membrane into cups. Continued MRCKβ activity then drives recruitment of a mechanosensing bridge enabling transmission of the cytoskeletal force required for cup closure and particle internalization. MRCKβ is also required for Fc receptor-mediated phagocytosis by macrophages. In vivo, MRCKβ is essential for RPE phagocytosis of photoreceptor debris and, hence, retinal integrity. MerTK-independent activation of MRCKβ signaling by a phosphomimetic Dbl3 mutant rescues phagocytosis in retinitis pigmentosa RPE cells lacking functional MerTK. Thus, conserved MRCKβ signaling controls spatiotemporal regulation of actomyosin contractility to guide actomyosin dynamics-driven phagocytosis and represents the principle phagocytic effector pathway downstream of MerTK.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128125784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Atg39 selectively captures inner nuclear membrane into lumenal vesicles for delivery to the autophagosome Atg39选择性地捕获核膜进入管腔小泡，并将其传递给自噬体

The Journal of Cell Biology Pub Date : 2021-02-22 DOI: 10.1101/2021.02.22.432332

Sunandini Chandra, Philip J. Mannino, David J. Thaller, Nicholas R. Ader, M. King, T. Melia, C. Lusk

引用次数: 14

The E3 ligase TRIM1 ubiquitinates LRRK2 and controls its localization, degradation, and toxicity E3连接酶TRIM1泛素化LRRK2并控制其定位、降解和毒性

The Journal of Cell Biology Pub Date : 2020-10-21 DOI: 10.1101/2020.10.21.336578

Adrienne E. D. Stormo, Farbod Shavarebi, Molly FitzGibbon, Elizabeth M. Earley, Hannah D. Ahrendt, Lotus Lum, E. Verschueren, D. Swaney, G. Skibinski, A. Ravisankar, J. van Haren, Emily J. Davis, Jeffrey R. Johnson, J. V. Dollen, Carson Balen, Jacob Porath, C. Crosio, C. Mirescu, C. Iaccarino, W. Dauer, R. Nichols, T. Wittmann, T. Cox, S. Finkbeiner, N. Krogan, S. Oakes, A. Hiniker

{"title":"The E3 ligase TRIM1 ubiquitinates LRRK2 and controls its localization, degradation, and toxicity","authors":"Adrienne E. D. Stormo, Farbod Shavarebi, Molly FitzGibbon, Elizabeth M. Earley, Hannah D. Ahrendt, Lotus Lum, E. Verschueren, D. Swaney, G. Skibinski, A. Ravisankar, J. van Haren, Emily J. Davis, Jeffrey R. Johnson, J. V. Dollen, Carson Balen, Jacob Porath, C. Crosio, C. Mirescu, C. Iaccarino, W. Dauer, R. Nichols, T. Wittmann, T. Cox, S. Finkbeiner, N. Krogan, S. Oakes, A. Hiniker","doi":"10.1101/2020.10.21.336578","DOIUrl":"https://doi.org/10.1101/2020.10.21.336578","url":null,"abstract":"Missense mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson’s Disease (PD); however, pathways regulating LRRK2 subcellular localization, function, and turnover are not fully defined. We performed quantitative mass spectrometry-based interactome studies to identify 48 novel LRRK2 interactors, including the microtubule-associated E3 ubiquitin ligase TRIM1 (Tripartite Motif Family 1). TRIM1 recruits LRRK2 to the microtubule cytoskeleton for ubiquitination and proteasomal degradation by binding LRRK2911-920, a nine amino acid segment within a flexible interdomain region (LRRK2853-981), which we designate the “Regulatory Loop” (RL). Phosphorylation of LRRK2 Ser910/Ser935 within LRRK2 RL serves as a molecular switch controlling LRRK2’s association with cytoplasmic 14-3-3 versus microtubule-bound TRIM1. Association with TRIM1 modulates LRRK2’s interaction with Rab29 and prevents upregulation of LRRK2 kinase activity by Rab29 in an E3-ligase-dependent manner. Finally, TRIM1 rescues neurite outgrowth deficits caused by PD-driving mutant LRRK2 G2019S. Our data suggest that TRIM1 is a critical regulator of LRRK2, controlling its degradation, localization, binding partners, kinase activity, and cytotoxicity.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"46 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114962629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Pathogenic mutations in the kinesin-3 motor KIF1A diminish force generation and movement through allosteric mechanisms 驱动蛋白-3运动KIF1A的致病性突变通过变构机制减少力的产生和运动

The Journal of Cell Biology Pub Date : 2020-09-03 DOI: 10.1101/2020.09.03.281576

B. Budaitis, S. Jariwala, Lu Rao, D. Sept, K. Verhey, A. Gennerich

{"title":"Pathogenic mutations in the kinesin-3 motor KIF1A diminish force generation and movement through allosteric mechanisms","authors":"B. Budaitis, S. Jariwala, Lu Rao, D. Sept, K. Verhey, A. Gennerich","doi":"10.1101/2020.09.03.281576","DOIUrl":"https://doi.org/10.1101/2020.09.03.281576","url":null,"abstract":"The kinesin-3 motor KIF1A functions in neurons where its fast and superprocessive motility is thought to be critical for long-distance transport. However, little is known about the force-generating properties of kinesin-3 motors. Using optical tweezers, we demonstrate that KIF1A and its C. elegans homolog UNC-104 undergo force-dependent detachments at ~3 pN and then rapidly reattach to the microtubule to resume motion, resulting in a sawtooth pattern of clustered force generation events that is unique among the kinesin superfamily. Whereas UNC-104 motors stall before detaching, KIF1A motors do not. To examine the mechanism of KIF1A force generation, we introduced mutations linked to human neurodevelopmental disorders, V8M and Y89D, based on their location in structural elements required for force generation in kinesin-1. Molecular dynamics simulations predict that the V8M and Y89D mutations impair docking of the N-terminal (β9) or C-terminal (β10) portions of the neck linker, respectively, to the KIF1A motor domain. Indeed, both mutations dramatically impair force generation of KIF1A but not the motor’s ability to rapidly reattach to the microtubule track. Homodimeric and heterodimeric mutant motors also display decreased velocities, run lengths, and landing rates and homodimeric Y89D motors exhibit a higher frequency of non-productive, diffusive events along the microtubule. In cells, cargo transport by the mutant motors is delayed. Our work demonstrates the importance of the neck linker in the force generation of kinesin-3 motors and advances our understanding of how mutations in the kinesin motor domain can manifest in disease.","PeriodicalId":343306,"journal":{"name":"The Journal of Cell Biology","volume":"18 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122271577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 39