Cytoskeleton (Hoboken, N.J.)最新文献_第9页

Conformational Regulation of Vertebrate γ-Tubulin Ring Complexes by CM1 Proteins. CM1蛋白对脊椎动物γ-微管蛋白环复合物的构象调控

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-08-01 Epub Date: 2024-12-18 DOI: 10.1002/cm.21979

Michal Wieczorek

引用次数: 0

Astral Microtubules Are Dispensable for Pavarotti Localization During Drosophila Spermatogonial Mitoses. 在果蝇精原细胞有丝分裂过程中，星体微管对帕瓦罗蒂定位是必不可少的。

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-08-01 Epub Date: 2025-01-03 DOI: 10.1002/cm.21986

Maria Giovanna Riparbelli, Massimo Migliorini, Giuliano Callaini

{"title":"Astral Microtubules Are Dispensable for Pavarotti Localization During Drosophila Spermatogonial Mitoses.","authors":"Maria Giovanna Riparbelli, Massimo Migliorini, Giuliano Callaini","doi":"10.1002/cm.21986","DOIUrl":"10.1002/cm.21986","url":null,"abstract":"We analysed here the dynamic of the kinesin-like Pavarotti (Pav) during male gametogenesis of wild-type and Sas4 mutant flies. Pav localizes to the equatorial region and the inner central spindle of late anaphase wild-type spermatogonia and displays a strong concentration at the midbody during late telophase. At metaphase of the first meiotic division, Pav shows widespread localization on the equatorial region of the spermatocytes. This unusual distribution restricts and enhances during anaphase where antiparallel cortical microtubules overlap. Additional Pav staining is also found in the inner central spindle where the microtubules overlap between the segregating chromosomes. At late telophase, Pav accumulates to the midbody and on a weak ring that surround the cytoplasmic bridges. Pav localizes in an equatorial discontinuous ring of Sas4 spermatogonia where the non-centrosomal microtubules overlap, but the motor protein is absent in the interior central spindle where the inner microtubules are lacking. However, the anastral spindles properly support cell division, suggesting that astral microtubules are dispensable for Pav localization in the Sas4 spermatogonial cell cortex. This function is presumably replaced by the antiparallel cortical microtubules extending from the acentriolar polar regions. In contrast, the majority of the meiotic spindles in Sas4 mutant testes do not progress beyond late anaphase, and only a small fraction of the primary spermatocytes experienced an abnormal division with the assembly of aberrant telophase spindles. Pav accumulates around the chromatin clusters or enhanced at the plus ends of the antiparallel non-centrosomal cortical bundles of microtubules. However, these bundles are not arranged properly in the equatorial region of the cell and cytokinesis is abnormal or fails. Therefore, the observations in Sas4 mutant testes suggest that the spermatogonial mitoses correctly occur in the absence of astral microtubules, whereas meiotic divisions fail.","PeriodicalId":72766,"journal":{"name":"Cytoskeleton (Hoboken, N.J.)","volume":" ","pages":"516-528"},"PeriodicalIF":1.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928763","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: 0

Author Profile: Khanh Pham. 作者简介：Khanh Pham。

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-08-01 Epub Date: 2024-12-24 DOI: 10.1002/cm.21981

Khanh Pham

引用次数: 0

Recombinant Monoclonal Antibodies for Detecting the Tubulin Post-Translational Modifications Glutamylation and Lysine-40 Acetylation. 检测微管蛋白翻译后修饰谷氨酰化和赖氨酸-40乙酰化的重组单克隆抗体。

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-07-12 DOI: 10.1002/cm.70011

Lynne Blasius, Ezekiel C Thomas, Erik H Collet, Erin Jenson, Chad G Pearson, Ryoma Ohi, Kristen J Verhey

{"title":"Recombinant Monoclonal Antibodies for Detecting the Tubulin Post-Translational Modifications Glutamylation and Lysine-40 Acetylation.","authors":"Lynne Blasius, Ezekiel C Thomas, Erik H Collet, Erin Jenson, Chad G Pearson, Ryoma Ohi, Kristen J Verhey","doi":"10.1002/cm.70011","DOIUrl":"10.1002/cm.70011","url":null,"abstract":"Post-translational modifications (PTMs) to tubulin subunits in microtubule filaments are thought to comprise a component of the tubulin code that specifies microtubule functions in cell physiology and animal development. Acetylation of Lysine-40 (K40) on α-tubulin (αTub-K40ac) and glutamylation of both α- and β-tubulin are two tubulin PTMs of interest to the field. Antibodies that recognize these PTMs have been indispensable tools to study the localization of these PTMs as well as their biological functions. Although widely used, these antibodies are procured from commercial sources and thus have drawbacks including availability, high cost, and lack of reproducibility. To mitigate these downsides, we report the protein sequences of GT335 (anti-glutamylation) and 6-11B-1 (anti-αTub-K40ac) monoclonal antibodies and describe the use of these sequences to generate recombinant monoclonal antibody (rMAb) versions of GT335 and 6-11B-1. We demonstrate through western blotting and immunofluorescence of cultured mammalian cells and Tetrahymena thermophila that rMAb-GT335 and rMAb-611B1 match the specific activity of the commercially available antibodies. Our work provides the field with a renewable source of antibodies with high specificity and affinity towards tubulin glutamylation and acetylation and opens the door to more reproducible and large-scale studies of the function and regulation these tubulin PTMs.","PeriodicalId":72766,"journal":{"name":"Cytoskeleton (Hoboken, N.J.)","volume":" ","pages":""},"PeriodicalIF":1.6,"publicationDate":"2025-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12276849/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144621506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Purification, Fluorescent Labeling, and Detyrosination of Mammalian Cell Tubulin for Biochemical Assays. 哺乳动物细胞微管蛋白的纯化、荧光标记和去酪氨酸的生化分析。

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-07-12 DOI: 10.1002/cm.70005

Ezekiel C Thomas, Yang Yue, Morgan L Pimm, Takashi Hotta, Ryoma Ohi, Kristen J Verhey

{"title":"Purification, Fluorescent Labeling, and Detyrosination of Mammalian Cell Tubulin for Biochemical Assays.","authors":"Ezekiel C Thomas, Yang Yue, Morgan L Pimm, Takashi Hotta, Ryoma Ohi, Kristen J Verhey","doi":"10.1002/cm.70005","DOIUrl":"10.1002/cm.70005","url":null,"abstract":"Microtubules play essential roles in numerous cellular processes. All microtubules are built from the protein tubulin, yet individual microtubules can differ spatially and temporally due to their tubulin isotype composition and post-translational modifications (PTMs). The tubulin code hypothesis posits that these differences can regulate microtubule function. However, investigating the properties of specific tubulin PTMs in vitro has been challenging because most reconstitution assays rely on tubulin purified from brain tissue that contains highly heterogeneous and modified microtubules. In this study, we present an optimized method for the purification of milligram quantities of unmodified tubulin from large-scale cultures of HeLa S3 cells. We also describe steps for efficient chemical labeling of tubulin and the generation of controlled tubulin PTMs. These tubulins can be used in microscopy or biochemistry-based experiments to investigate how the tubulin code influences microtubule properties and functions. Overall, our method is easily adaptable, highly reproducible, and broadly accessible to labs with general equipment.","PeriodicalId":72766,"journal":{"name":"Cytoskeleton (Hoboken, N.J.)","volume":" ","pages":""},"PeriodicalIF":1.6,"publicationDate":"2025-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12276855/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144621505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Toxoplasma replication is inhibited by MMV676477 without development of resistance. MMV676477 可抑制弓形虫的复制，但不会产生抗药性。

IF 1.6

Cytoskeleton (Hoboken, N.J.) Pub Date : 2025-01-01 Epub Date: 2024-05-16 DOI: 10.1002/cm.21876

Izra Abbaali, Danny Truong, Dawn M Wetzel, Naomi S Morrissette

{"title":"Toxoplasma replication is inhibited by MMV676477 without development of resistance.","authors":"Izra Abbaali, Danny Truong, Dawn M Wetzel, Naomi S Morrissette","doi":"10.1002/cm.21876","DOIUrl":"10.1002/cm.21876","url":null,"abstract":"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 Leishmania amazonensis and Trypanosoma brucei, and the apicomplexan parasite Plasmodium falciparum by selectively stabilizing protozoan microtubules. In this report, we show that MMV676477 inhibits intracellular growth of the human apicomplexan pathogen Toxoplasma gondii 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 Toxoplasma 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.","PeriodicalId":72766,"journal":{"name":"Cytoskeleton (Hoboken, N.J.)","volume":" ","pages":"5-11"},"PeriodicalIF":1.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11568068/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140960164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An interview with Peter Gunning - School of Medical Sciences, UNSW Sydney, Australia. 采访彼得·甘宁-医学科学学院，新南威尔士大学悉尼，澳大利亚。

Cytoskeleton (Hoboken, N.J.) Pub Date : 2022-04-01 Epub Date: 2022-07-22 DOI: 10.1002/cm.21715

Paul Trevorrow, Peter Gunning

引用次数: 0

Meet the editorial staff: Interview series. 与编辑人员见面：访谈系列。

Cytoskeleton (Hoboken, N.J.) Pub Date : 2022-04-01 Epub Date: 2022-07-13 DOI: 10.1002/cm.21714

Paul Trevorrow

引用次数: 0

Follow that cell: Leukocyte migration in L-plastin mutant zebrafish. 跟随细胞:l -活蛋白突变斑马鱼的白细胞迁移。

Cytoskeleton (Hoboken, N.J.) Pub Date : 2022-04-01 Epub Date: 2022-07-22 DOI: 10.1002/cm.21717

John B Linehan, Jose Lucas Zepeda, Taylor A Mitchell, Elizabeth E LeClair

{"title":"Follow that cell: Leukocyte migration in L-plastin mutant zebrafish.","authors":"John B Linehan, Jose Lucas Zepeda, Taylor A Mitchell, Elizabeth E LeClair","doi":"10.1002/cm.21717","DOIUrl":"https://doi.org/10.1002/cm.21717","url":null,"abstract":"Actin assemblies are important in motile cells such as leukocytes which form dynamic plasma membrane extensions or podia. L-plastin (LCP1) is a leukocyte-specific calcium-dependent actin-bundling protein that, in mammals, is known to affect immune cell migration. Previously, we generated CRISPR/Cas9 engineered zebrafish lacking L-plastin (lcp1-/-) and reported that they had reduced survival to adulthood, suggesting that lack of L-plastin might negatively affect the immune system. To test this hypothesis, we examined the distribution and migration of neutrophils and macrophages in the transparent tail of early zebrafish larvae using cell-specific markers and an established wound-migration assay. Knockout larvae were similar to their heterozygous siblings in having equal body sizes and comparable numbers of neutrophils in caudal hematopoietic tissue at two days post-fertilization, indicating no gross defect in neutrophil production or developmental migration. When stimulated by a tail wound, all genotypes of neutrophils were equally migratory in a two-hour window. However for macrophages we observed both migration defects and morphological differences. L-plastin knockout macrophages still homed to wounds but were slower, less directional and had a star-like morphology with many leading and trailing projections. In contrast, wild type macrophages were faster, more directional, and had a more streamlined, slug-like morphology. Overall, these findings show that in larval zebrafish L-plastin knockout primarily affects the macrophage response with possible consequences for organismal immunity. Consistent with our observations, we propose a model in which cytoplasmic L-plastin negatively regulates macrophage integrin adhesion by holding these transmembrane heterodimers in a ‘clasped’, inactive form and is a necessary part of establishing macrophage polarity during chemokine-induced motility.","PeriodicalId":72766,"journal":{"name":"Cytoskeleton (Hoboken, N.J.)","volume":"79 4-5","pages":"26-37"},"PeriodicalIF":0.0,"publicationDate":"2022-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40488845","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: 1

Cytoskeleton: Methods and Protocols 细胞骨架:方法和方案

Cytoskeleton (Hoboken, N.J.) Pub Date : 2022-01-01 DOI: 10.1007/978-1-0716-1661-1

Marina Daecher

引用次数: 0