Cytoskeleton最新文献

{"title":"A comparative analysis of paxillin and Hic-5 proximity interactomes","authors":"Katia Brock,&nbsp;Kyle M. Alpha,&nbsp;Grant Brennan,&nbsp;Ebbing P. De Jong,&nbsp;Elizabeth Luke,&nbsp;Christopher E. Turner","doi":"10.1002/cm.21878","DOIUrl":"10.1002/cm.21878","url":null,"abstract":"<p>Focal adhesions serve as structural and signaling hubs, facilitating bidirectional communication at the cell–extracellular matrix interface. Paxillin and the related Hic-5 (TGFβ1i1) are adaptor/scaffold proteins that recruit numerous structural and regulatory proteins to focal adhesions, where they perform both overlapping and discrete functions. In this study, paxillin and Hic-5 were expressed in U2OS osteosarcoma cells as biotin ligase (BioID2) fusion proteins and used as bait proteins for proximity-dependent biotinylation in order to directly compare their respective interactomes. The fusion proteins localized to both focal adhesions and the centrosome, resulting in biotinylation of components of each of these structures. Biotinylated proteins were purified and analyzed by mass spectrometry. The list of proximity interactors for paxillin and Hic-5 comprised numerous shared core focal adhesion proteins that likely contribute to their similar functions in cell adhesion and migration, as well as proteins unique to paxillin and Hic-5 that have been previously localized to focal adhesions, the centrosome, or the nucleus. Western blotting confirmed biotinylation and enrichment of FAK and vinculin, known interactors of Hic-5 and paxillin, as well as several potentially unique proximity interactors of Hic-5 and paxillin, including septin 7 and ponsin, respectively. Further investigation into the functional relationship between the unique interactors and Hic-5 or paxillin may yield novel insights into their distinct roles in cell migration.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"82 1-2","pages":"12-31"},"PeriodicalIF":2.4,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141155321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The mechanics of cilia and flagella: What we know and what we need to know","authors":"Charles B. Lindemann,&nbsp;Kathleen A. Lesich","doi":"10.1002/cm.21879","DOIUrl":"10.1002/cm.21879","url":null,"abstract":"<p>In this review, we provide a condensed overview of what is currently known about the mechanical functioning of the flagellar/ciliary axoneme. We also present a list of 10 specific areas where our current knowledge is incomplete and explain the benefits of further experimental investigation. Many of the physical parameters of the axoneme and its component parts have not been determined. This limits our ability to understand how the axoneme structure contributes to its functioning in several regards. It restricts our ability to understand how the mechanics of the structure contribute to the regulation of motor function. It also confines our ability to understand the three-dimensional workings of the axoneme and how various beating modes are accomplished. Lastly, it prevents accurate computational modeling of the axoneme in three-dimensions.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 11","pages":"648-668"},"PeriodicalIF":2.4,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21879","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141082622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vascular endothelial cell morphology and alignment regulate VEGF-induced endothelial nitric oxide synthase activation","authors":"Aparna Bhattacharyya,&nbsp;Kenneth A. Barbee","doi":"10.1002/cm.21872","DOIUrl":"10.1002/cm.21872","url":null,"abstract":"<p>Nitric oxide (NO) production by endothelial nitric oxide synthase (eNOS) inhibits platelet and leukocyte adhesion while promoting vasorelaxation in smooth muscle cells. Dysfunctional regulation of eNOS is a hallmark of various vascular pathologies, notably atherosclerosis, often associated with areas of low shear stress on endothelial cells (ECs). While the link between EC morphology and local hemodynamics is acknowledged, the specific impact of EC morphology on eNOS regulation remains unclear. Morphological differences between elongated, aligned ECs and polygonal, randomly oriented ECs correspond to variations in focal adhesion and cytoskeletal organization, suggesting differing levels of cytoskeletal prestress. However, the functional outcomes of cytoskeletal prestress, particularly in the absence of shear stress, are not extensively studied in ECs. Some evidence suggests that elongated ECs exhibit decreased immunogenicity and enhanced NO production. This study aims to elucidate the signaling pathways governing VEGF-stimulated eNOS regulation in the aligned EC phenotype characterized by elongated and aligned cells within a monolayer. Using anisotropic topographic cues, bovine aortic endothelial cells (BAECs) were elongated and aligned, followed by VEGF treatment in the presence or absence of cytoskeletal tension inhibitors. Phosphorylation of eNOS ser1179, AKT ser437 and FAK Tyr397 in response to VEGF challenge were significantly heightened in aligned ECs compared to unaligned ECs. Moreover this response proved to be robustly tied to cytoskeletal tension as evinced by the abrogation of responses in the presence of the myosin II ATPase inhibitor, blebbistatin. Notably, this work demonstrates for the first time the reliance on FAK phosphorylation in VEGF-mediated eNOS activation and the comparatively greater contribution of the cytoskeletal machinery in propagating VEGF-eNOS signaling in aligned and elongated ECs. This research underscores the importance of utilizing appropriate vascular models in drug development and sheds light on potential mechanisms underlying vascular function and pathology that can help inform vascular graft design.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 9-10","pages":"473-487"},"PeriodicalIF":2.4,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141075693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Growth-induced collective bending and kinetic trapping of cytoskeletal filaments","authors":"Deb Sankar Banerjee,&nbsp;Simon L. Freedman,&nbsp;Michael P. Murrell,&nbsp;Shiladitya Banerjee","doi":"10.1002/cm.21877","DOIUrl":"10.1002/cm.21877","url":null,"abstract":"<p>Growth and turnover of actin filaments play a crucial role in the construction and maintenance of actin networks within cells. Actin filament growth occurs within limited space and finite subunit resources in the actin cortex. To understand how filament growth shapes the emergent architecture of actin networks, we developed a minimal agent-based model coupling filament mechanics and growth in a limiting subunit pool. We find that rapid filament growth induces kinetic trapping of highly bent actin filaments. Such collective bending patterns are long-lived, organized around nematic defects, and arise from competition between filament polymerization and bending elasticity. The stability of nematic defects and the extent of kinetic trapping are amplified by an increase in the abundance of the actin pool and by crosslinking the network. These findings suggest that kinetic trapping is a robust consequence of growth in crowded environments, providing a route to program shape memory in actin networks.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"409-419"},"PeriodicalIF":2.4,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21877","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141075705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TNF-alpha promotes cilia elongation via mixed lineage kinases signaling in mouse fibroblasts and human RPE-1 cells","authors":"Amrita Kumari,&nbsp;Amada D. Caliz,&nbsp;Hyung-Jin Yoo,&nbsp;Shashi Kant,&nbsp;Anastassiia Vertii","doi":"10.1002/cm.21873","DOIUrl":"10.1002/cm.21873","url":null,"abstract":"<p>The primary cilium is a characteristic feature of most non-immune cells and functions as an environmental signal transduction sensor. The defects in primary cilium have profound effects on the developmental program, including the maturation of retinal epithelium. The ciliary length is tightly regulated during ciliogenesis, but the impact of inflammation on ciliary length remains elusive. The current study investigates the outcome of inflammatory stimuli for the primary cilium length in retinal epithelium cells and mouse embryonic fibroblasts. Here, we report that exposure to the pro-inflammatory cytokine TNF-alpha elongates cilia in a mixed-lineage kinase (MLK)-dependent manner. Pro-inflammatory stimuli such as bacterial LPS and interferon-gamma have similar effects on ciliary length. In contrast, febrile condition-mimicking heat stress dramatically reduced the number of ciliated cells regardless of TNF-alpha exposure but did not shorten TNF-induced elongation, suggesting distinct but rapid effects of inflammatory stresses on ciliogenesis.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 11","pages":"639-647"},"PeriodicalIF":2.4,"publicationDate":"2024-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141066212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nucleating amoeboid cancer cell motility with Diaphanous related formins","authors":"Neelakshi Kar,&nbsp;Jeremy S. Logue","doi":"10.1002/cm.21880","DOIUrl":"10.1002/cm.21880","url":null,"abstract":"<p>The tissue invasive capacity of cancer cells is determined by their phenotypic plasticity. For instance, mesenchymal to amoeboid transition has been found to facilitate the passage of cancer cells through confined environments. This phenotypic transition is also heavily regulated by the architecture of the actin cytoskeleton, which may increase myosin contractility and the intracellular pressure that is known to drive bleb formation. In this review, we highlight several Diaphanous related formins (DRFs) that have been found to promote or suppress bleb formation in cancer cells, which is a hallmark of amoeboid migration. Based on the work discussed here, the role of the DRFs in cancer(s) is worthy of further scrutiny in animal models, as they may prove to be therapeutic targets.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"82 3","pages":"91-97"},"PeriodicalIF":2.4,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11570701/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140960134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The unique properties of Big tau in the visual system","authors":"Itzhak Fischer,&nbsp;Theresa Connors,&nbsp;Julien Bouyer,&nbsp;Ying Jin","doi":"10.1002/cm.21875","DOIUrl":"10.1002/cm.21875","url":null,"abstract":"<p>Tau is a microtubule associated protein that plays important roles in regulating the properties of microtubules and axonal transport, as well as tauopathies associated with toxic aggregates leading to neurodegenerative diseases. It is encoded by the MAPT gene forming multiple isoforms (45–60 kDa) by alternative splicing which are developmentally regulated. The high molecular weight (MW) tau isoform of 105 kDa, termed Big tau, was originally discovered in the peripheral nervous system (PNS) but later found in selective CNS areas. It contains an additional large exon 4a generating a long projecting domain of about 250 amino acids. Here we investigated the properties of Big tau in the visual system of rats, its distribution in retinal ganglion cells and the optic nerve as well as its developmental regulation using biochemical, molecular and histological analyses. We discovered that Big tau is expresses as a 95 kDa protein (termed middle MW) containing exons 4a, 6 as well as exon 10 which defines a 4 microtubule-binding repeats (4R). It lacks exons 2/3 but shares the extensive phosphorylation characteristic of other tau isoforms. Importantly, early in development the visual system expresses only the low MW isoform (3R) switching to both the low and middle MW isoforms (4R) in adult retinal ganglion neurons and their corresponding axons. This is a unique structure and expression pattern of Big tau, which we hypothesize is associated with the specific properties of the visual system different from what has been previously described in the PNS and other areas of the nervous system.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 9-10","pages":"488-499"},"PeriodicalIF":2.4,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140960163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Author profile—Prashali Chauhan","authors":"Prashali Chauhan","doi":"10.1002/cm.21874","DOIUrl":"10.1002/cm.21874","url":null,"abstract":"","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"420-421"},"PeriodicalIF":2.4,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140960068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Toxoplasma replication is inhibited by MMV676477 without development of resistance","authors":"Izra Abbaali,&nbsp;Danny Truong,&nbsp;Dawn M. Wetzel,&nbsp;Naomi S. Morrissette","doi":"10.1002/cm.21876","DOIUrl":"10.1002/cm.21876","url":null,"abstract":"<p>Protozoan parasites cause life-threatening infections in both humans and animals, including agriculturally significant livestock. Available treatments are typically narrow spectrum and are complicated by drug toxicity and the development of resistant parasites. Protozoan tubulin is an attractive target for the development of broad-spectrum antimitotic agents. The Medicines for Malaria Pathogen Box compound MMV676477 was previously shown to inhibit replication of kinetoplastid parasites, such as <i>Leishmania amazonensis</i> and <i>Trypanosoma brucei</i>, and the apicomplexan parasite <i>Plasmodium falciparum</i> by selectively stabilizing protozoan microtubules. In this report, we show that MMV676477 inhibits intracellular growth of the human apicomplexan pathogen <i>Toxoplasma gondii</i> with an EC₅₀ value of ~50 nM. MMV676477 does not stabilize vertebrate microtubules or cause other toxic effects in human fibroblasts. The availability of tools for genetic studies makes <i>Toxoplasma</i> a useful model for studies of the cytoskeleton. We conducted a forward genetics screen for MMV676477 resistance, anticipating that missense mutations would delineate the binding site on protozoan tubulin. Unfortunately, we were unable to use genetics to dissect target interactions because no resistant parasites emerged. This outcome suggests that future drugs based on the MMV676477 scaffold would be less likely to be undermined by the emergence of drug resistance.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"82 1-2","pages":"5-11"},"PeriodicalIF":2.4,"publicationDate":"2024-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21876","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140969849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrastructure expansion microscopy (U-ExM) of mouse and human kidneys for analysis of subcellular structures","authors":"Ewa Langner,&nbsp;Pongpratch Puapatanakul,&nbsp;Rachel Pudlowski,&nbsp;Dema Yaseen Alsabbagh,&nbsp;Jeffrey H. Miner,&nbsp;Amjad Horani,&nbsp;Susan K. Dutcher,&nbsp;Steven L. Brody,&nbsp;Jennifer T. Wang,&nbsp;Hani Y. Suleiman,&nbsp;Moe R. Mahjoub","doi":"10.1002/cm.21870","DOIUrl":"10.1002/cm.21870","url":null,"abstract":"<p>Ultrastructure expansion microscopy (U-ExM) involves the physical magnification of specimens embedded in hydrogels, which allows for super-resolution imaging of subcellular structures using a conventional diffraction-limited microscope. Methods for expansion microscopy exist for several organisms, organs, and cell types, and used to analyze cellular organelles and substructures in nanoscale resolution. Here, we describe a simple step-by-step U-ExM protocol for the expansion, immunostaining, imaging, and analysis of cytoskeletal and organellar structures in kidney tissue. We detail the critical modified steps to optimize isotropic kidney tissue expansion, and preservation of the renal cell structures of interest. We demonstrate the utility of the approach using several markers of renal cell types, centrioles, cilia, the extracellular matrix, and other cytoskeletal elements. Finally, we show that the approach works well on mouse and human kidney samples that were preserved using different fixation and embedding conditions. Overall, this protocol provides a simple and cost-effective approach to analyze both preclinical and clinical renal samples in high detail, using conventional lab supplies and standard widefield or confocal microscopy.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 11","pages":"618-638"},"PeriodicalIF":2.4,"publicationDate":"2024-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140878074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}