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https://www.readbyqxmd.com/read/28643946/activated-proline-rich-tyrosine-kinase-2-regulates-meiotic-spindle-assembly-in-the-mouse-oocyte
#1
Xiao-Qian Meng, Bing Cui, Dong Cheng, Hui Lyu, Li-Gang Jiang, Ke-Gang Zheng, Shu-Zhen Liu, Jie Pan, Cong Zhang, Jing Bai, Jun Zhou
Proline-rich tyrosine kinase 2 (PYK2), a member of the protein tyrosine kinase family, plays an important role in various cellular processes. PYK2 can be phosphorylated on tyrosine 402 by diverse stimuli at the cell surface, and recent studies have shown that this activated form of PYK2 is enriched in oocytes and required for fertilization. However, the subcellular localization and functions of activated PYK2 in oocytes remain elusive. In this study, we demonstrate that the localization of p-PYK2 undergoes dynamic changes during in vitro maturation of mouse oocytes...
June 23, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28641615/-expression-and-asymmetric-division-of-numb-in-leukemia-cell-k562-line
#2
Zheng Li, Huan Li, Yi-Hui Li, Ying-Xi Xu, Hai-Yan Xing, Ke-Jing Tang, Zheng Tian, Min Wang, Qing Rao
OBJECTIVE: To investigate the role of asymmetric division in leukemia cells through detection of expression and asymmetric division of Numb in differentiated and undifferentiated K562 cells. METHODS: Firstly, Hemin was used to induce K562 cell differentiation, and the expression of Numb was detected by the real-time quantitative RT-PCR and flow cytometry. After K562 cells were synchronized by nocodazole, the Numb protein was labeled by immunohistochemical staining, followed by the determination of the terminally differentiated cells through confocal microscopy...
June 2017: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28640209/quinolin-6-yloxyacetamides-are-microtubule-destabilizing-agents-that-bind-to-the-colchicine-site-of-tubulin
#3
Ashwani Sharma, Gonzalo Sáez-Calvo, Natacha Olieric, Francisco de Asís Balaguer, Isabel Barasoain, Clemens Lamberth, J Fernando Díaz, Michel O Steinmetz
Quinolin-6-yloxyacetamides (QAs) are a chemical class of tubulin polymerization inhibitors that were initially identified as fungicides. Here, we report that QAs are potent anti-proliferative agents against human cancer cells including ones that are drug-resistant. QAs act by disrupting the microtubule cytoskeleton and by causing severe mitotic defects. We further demonstrate that QAs inhibit tubulin polymerization in vitro. The high resolution crystal structure of the tubulin-QA complex revealed that QAs bind to the colchicine site on tubulin, which is targeted by microtubule-destabilizing agents such as colchicine and nocodazole...
June 22, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28576942/wild-type-monomeric-%C3%AE-synuclein-can-impair-vesicle-endocytosis-and-synaptic-fidelity-via-tubulin-polymerization-at-the-calyx-of-held
#4
Kohgaku Eguchi, Zacharie Taoufiq, Oliver Thorn-Seshold, Dirk Trauner, Masato Hasegawa, Tomoyuki Takahashi
α-Synuclein is a presynaptic protein, the function of which has as yet to be identified, but its neuronal content increases in patients of synucleinopathies including Parkinson's disease. Chronic overexpression of α-synuclein reportedly expresses various phenotypes of synaptic dysfunction, but the primary target of its toxicity has not been determined. To investigate this, we acutely loaded human recombinant α-synuclein or its pathological mutants in their monomeric forms into the calyces of Held presynaptic terminals in slices from auditorily mature and immature rats of either sexes...
June 2, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28562161/pp2a-b56%C3%AE-is-required-for-an-efficient-spindle-assembly-checkpoint
#5
Prajakta Varadkar, Fatima Abbasi, Kazuyo Takeda, Jade J Dyson, Brent McCright
The Spindle Assembly Checkpoint (SAC) is part of a complex feedback system designed to ensure that cells do not proceed through mitosis unless all chromosomal kinetochores have attached to spindle microtubules. The formation of the kinetochore complex and the implementation of the SAC are regulated by multiple kinases and phosphatases. BubR1 is a phosphoprotein that is part of the Cdc20 containing mitotic checkpoint complex that inhibits the APC/C so that Cyclin B1 and Securin are not degraded, thus preventing cells going into anaphase...
May 31, 2017: Cell Cycle
https://www.readbyqxmd.com/read/28557611/lamin-a-and-microtubules-collaborate-to-maintain-nuclear-morphology
#6
Zeshan Tariq, Haoyue Zhang, Alexander Chia-Liu, Yang Shen, Yantenew Gete, Zheng-Mei Xiong, Claire Tocheny, Leonard Campanello, Di Wu, Wolfgang Losert, Kan Cao
Lamin A (LA) is a critical structural component of the nuclear lamina. Mutations within the LA gene (LMNA) lead to several human disorders, most striking of which is Hutchinson-Gilford Progeria Syndrome (HGPS), a premature aging disorder. HGPS cells are best characterized by an abnormal nuclear morphology known as nuclear blebbing, which arises due to the accumulation of progerin, a dominant mutant form of LA. The microtubule (MT) network is known to mediate changes in nuclear morphology in the context of specific events such as mitosis, cell polarization, nucleus positioning and cellular migration...
May 30, 2017: Nucleus
https://www.readbyqxmd.com/read/28548701/microtubules-regulate-brush-border-formation
#7
Facundo M Tonucci, Anabela Ferretti, Evangelina Almada, Pamela Cribb, Rodrigo Vena, Florencia Hidalgo, Cristián Favre, Matt J Tyska, Irina Kaverina, M Cecilia Larocca
Most epithelial cells contain apical membrane structures associated to bundles of actin filaments, which constitute the brush border. Whereas microtubule participation in the maintenance of the brush border identity has been characterized, their contribution to de novo microvilli organization remained elusive. Hereby, using a cell model of individual enterocyte polarization, we found that nocodazole induced microtubule depolymerization prevented the de novo brush border formation. Microtubule participation in brush border actin organization was confirmed in polarized kidney tubule MDCK cells...
May 26, 2017: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/28526408/semi-retentive-cytoskeletal-fractionation-sercyf-a-novel-method-for-the-biochemical-analysis-of-the-organization-of-microtubule-and-actin-cytoskeleton-networks
#8
Yuta Sato, Yota Murakami, Masayuki Takahashi
A variety of biochemical fractionation methods are available for the quantification of cytoskeletal components. However, each method is designed to target only one cytoskeletal network, either the microtubule (MT) or actin cytoskeleton, and non-targeted cytoskeletal networks are ignored. Considering the importance of MT-actin crosstalk, the organization of both the targeted and non-targeted cytoskeletal networks must be retained intact during fractionation for the accurate analysis of cytoskeletal organization...
May 17, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28428427/microtubules-acquire-resistance-from-mechanical-breakage-through-intralumenal-acetylation
#9
Zhenjie Xu, Laura Schaedel, Didier Portran, Andrea Aguilar, Jérémie Gaillard, M Peter Marinkovich, Manuel Théry, Maxence V Nachury
Eukaryotic cells rely on long-lived microtubules for intracellular transport and as compression-bearing elements. We considered that long-lived microtubules are acetylated inside their lumen and that microtubule acetylation may modify microtubule mechanics. Here, we found that tubulin acetylation is required for the mechanical stabilization of long-lived microtubules in cells. Depletion of the tubulin acetyltransferase TAT1 led to a significant increase in the frequency of microtubule breakage. Nocodazole-resistant microtubules lost upon removal of acetylation were largely restored by either pharmacological or physical removal of compressive forces...
April 21, 2017: Science
https://www.readbyqxmd.com/read/28420732/modulation-of-tmem16a-channel-activity-by-the-von-willebrand-factor-type-a-vwa-domain-of-the-calcium-activated-chloride-channel-regulator-1-clca1
#10
Monica Sala-Rabanal, Zeynep Yurtsever, Kayla N Berry, Colin G Nichols, Tom J Brett
Calcium-activated chloride channels (CaCCs) are key players in transepithelial ion transport and fluid secretion, smooth muscle constriction, neuronal excitability, and cell proliferation. The CaCC regulator 1 (CLCA1) modulates the activity of the CaCC TMEM16A/Anoctamin 1 (ANO1) by directly engaging the channel at the cell surface, but the exact mechanism is unknown. Here we demonstrate that the von Willebrand factor type A (VWA) domain within the cleaved CLCA1 N-terminal fragment is necessary and sufficient for this interaction...
June 2, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28413120/endosome-to-trans-golgi-network-transport-of-proprotein-convertase-7-is-mediated-by-a-cluster-of-basic-amino-acids-and-palmitoylated-cysteines
#11
Jeroen Declercq, Bruno Ramos-Molina, Ragna Sannerud, Bas Brouwers, Vincent P E G Pruniau, Sandra Meulemans, Evelyn Plets, Wim Annaert, John W M Creemers
Proprotein Convertase 7 (PC7) is a Furin-like endoprotease that cleaves precursor proteins at basic amino acids. PC7 is concentrated in the trans-Golgi network (TGN) but it shuttles between the plasma membrane and the TGN depending on sequences in the cytoplasmic tail. A short region containing a three amino acids motif, P(724)-L(725)-C(726), is essential and sufficient for internalization of PC7 but not for TGN localization, which requires the additional presence of the juxtamembrane region. In this study we have investigated the contribution of a cluster of basic amino acids and two reversibly palmitoylated cysteine residues to endocytic trafficking...
April 7, 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28412508/methyl-5-1h-indol-3-yl-selanyl-1h-benzoimidazol-2-ylcarbamate-m-24-a-novel-tubulin-inhibitor-causes-g2-m-arrest-and-cell-apoptosis-by-disrupting-tubulin-polymerization-in-human-cervical-and-breast-cancer-cells
#12
Daiying Zuo, Xuewei Jiang, Mengting Han, Jiwei Shen, Binyue Lang, Qi Guan, Zhaoshi Bai, Chunming Han, Zengqiang Li, Weige Zhang, Yingliang Wu
Methyl 5-[(1H-indol-3-yl)selanyl]-1H-benzoimidazol-2-ylcarbamate (M-24) is a newly synthesized analogue of nocodazole by our group and has been found to be active for some cancer cells. However, its sensitivity to different cell lines and the underlying anticancer mechanism are still unclear. In this study, we proved that M-24 had strong time- and dose-dependent anti-proliferative effects on human cervical cancer HeLa cells and human breast carcinoma MCF-7 cells. We demonstrated that the growth inhibitory effects of M-24 in both cell lines were associated with microtubule depolymerization...
April 13, 2017: Toxicology in Vitro: An International Journal Published in Association with BIBRA
https://www.readbyqxmd.com/read/28397141/differentiation-dependent-rearrangements-of-actin-filaments-and-microtubules-hinder-apical-endocytosis-in-urothelial-cells
#13
Larisa Tratnjek, Rok Romih, Mateja Erdani Kreft
During differentiation, superficial urothelial cells (UCs) of the urinary bladder form the apical surface, which is almost entirely covered by urothelial plaques containing densely packed uroplakin particles. These urothelial plaques are the main structural components of the blood-urine permeability barrier in the urinary bladder. We have shown previously that endocytosis from the apical plasma membrane decreases during urothelial cell differentiation. Here, we investigated the role of actin filament and microtubule rearrangements in apical endocytosis of differentiating UCs cells using hyperplastic and normoplastic porcine urothelial models...
April 10, 2017: Histochemistry and Cell Biology
https://www.readbyqxmd.com/read/28379780/akap95-interacts-with-nucleoporin-tpr-in-mitosis-and-is-important-for-the-spindle-assembly-checkpoint
#14
Graciela López-Soop, Torunn Rønningen, Agnieszka Rogala, Nina Richartz, Heidi Kiil Blomhoff, Bernd Thiede, Philippe Collas, Thomas Küntziger
Faithful chromosome segregation during mitosis relies on a proofreading mechanism that monitors proper kinetochore-microtubule attachments. The spindle assembly checkpoint (SAC) is based on the concerted action of numerous components that maintain a repressive signal inhibiting transition into anaphase until all chromosomes are attached. Here we show that A-Kinase Anchoring Protein 95 (AKAP95) is necessary for proper SAC function. AKAP95-depleted HeLa cells show micronuclei formed from lagging chromosomes at mitosis...
May 19, 2017: Cell Cycle
https://www.readbyqxmd.com/read/28351621/critical-roles-of-astrin-in-the-mitosis-of-immature-rat-sertoli-cells
#15
Yuki Tochigi, Yuka Iwasaki, Masanori Sano, Hidenori Yasuda, Kentaro Katayama, Hiroetsu Suzuki
Male hypogonadism (hgn/hgn) rats show testicular hypoplasia accompanied by dysplastic development of seminiferous tubules due to loss-of-function mutation of the gene encoding Astrin, which is required for mitotic progression in the division cycle of HeLa cells. In the present study, we examined the cytological base leading to the decrease of Sertoli cells in hgn/hgn testes. In hgn/hgn testes on postnatal day 3, anti-phospho-histone H3 (Ser10) (pH3)-positive mitotic phase and TUNEL-positive apoptosis increased in GATA4-positive Sertoli cells...
May 13, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28344766/phosphorylation-of-lsd1-by-plk1-promotes-its-chromatin-release-during-mitosis
#16
Bin Peng, Ruifeng Shi, Weiwei Jiang, Yue-He Ding, Meng-Qiu Dong, Wei-Guo Zhu, Xingzhi Xu
BACKGROUND: Lysine-specific histone demethylase 1 (LSD1) modulates chromatin status through demethylation of H3K4 and H3K9. It has been demonstrated that LSD1 is hyperphosphorylated and dissociates from chromatin during mitosis. However, the molecular mechanism of LSD1 detachment is unknown. RESULTS: In this report, we found that polo-like kinase 1 (PLK1) directly interacted with LSD1 and phosphorylated LSD1 at Ser-126 . Nocodazole-induced metaphase arrest promoted release of LSD1 from chromatin, and the phosphorylation-defective mutant LSD1 (S126A) failed to dissociate from chromatin upon nocodazole treatment...
2017: Cell & Bioscience
https://www.readbyqxmd.com/read/28328318/toward-discovery-of-novel-microtubule-targeting-agents-a-snap-tag-based-high-content-screening-assay-for-the-analysis-of-microtubule-dynamics-and-cell-cycle-progression
#17
Nina Berges, Katharina Arens, Verena Kreusch, Rainer Fischer, Stefano Di Fiore
Microtubule targeting agents (MTAs) are used for the treatment of cancer. Novel MTAs could provide additional and beneficial therapeutic options. To improve the sensitivity and throughput of standard immunofluorescence assays for the characterization of MTAs, we used SNAP-tag technology to produce recombinant tubulin monomers. To visualize microtubule filaments, A549 cells transfected with SNAP-tubulin were stained with a membrane-permeable, SNAP-reactive dye. The treatment of SNAP-tubulin cells with stabilizing MTAs such as paclitaxel resulted in the formation of coarsely structured microtubule filaments, whereas depolymerizing MTAs such as nocodazole resulted in diffuse staining patterns in which the tubulin filaments were no longer distinguishable...
April 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28242250/enhanced-stability-of-microtubules-contributes-in-the-development-of-colchicine-resistance-in-mcf-7-cells
#18
Ankit Rai, Sonia Kapoor, Afsana Naaz, Manas Kumar Santra, Dulal Panda
Understanding the mechanism of resistance to tubulin-targeted anticancer drugs is important for improved chemotherapy. In this work, a colchicine-resistant MCF-7 cell line (MCF-7Col30) was generated by the gradual increment of colchicine treatment and the MCF-7Col30 showed ∼8-fold resistance towards colchicine. MCF-7Col30 cells showed ∼2.5-fold resistance against microtubule depolymerizing agents, vinblastine, and nocodazole. In contrast, it displayed more sensitivity towards paclitaxel, a microtubule-polymerizing agent...
May 15, 2017: Biochemical Pharmacology
https://www.readbyqxmd.com/read/28219395/efficient-precise-knockin-with-a-double-cut-hdr-donor-after-crispr-cas9-mediated-double-stranded-dna-cleavage
#19
Jian-Ping Zhang, Xiao-Lan Li, Guo-Hua Li, Wanqiu Chen, Cameron Arakaki, Gary D Botimer, David Baylink, Lu Zhang, Wei Wen, Ya-Wen Fu, Jing Xu, Noah Chun, Weiping Yuan, Tao Cheng, Xiao-Bing Zhang
BACKGROUND: Precise genome editing via homology-directed repair (HDR) after double-stranded DNA (dsDNA) cleavage facilitates functional genomic research and holds promise for gene therapy. However, HDR efficiency remains low in some cell types, including some of great research and clinical interest, such as human induced pluripotent stem cells (iPSCs). RESULTS: Here, we show that a double cut HDR donor, which is flanked by single guide RNA (sgRNA)-PAM sequences and is released after CRISPR/Cas9 cleavage, increases HDR efficiency by twofold to fivefold relative to circular plasmid donors at one genomic locus in 293 T cells and two distinct genomic loci in iPSCs...
February 20, 2017: Genome Biology
https://www.readbyqxmd.com/read/28202041/the-microtubule-plus-end-tracking-protein-tacc3-promotes-persistent-axon-outgrowth-and-mediates-responses-to-axon-guidance-signals-during-development
#20
Burcu Erdogan, Garrett M Cammarata, Eric J Lee, Benjamin C Pratt, Andrew F Francl, Erin L Rutherford, Laura Anne Lowery
BACKGROUND: Formation of precise neuronal connections requires proper axon guidance. Microtubules (MTs) of the growth cone provide a critical driving force during navigation of the growing ends of axons. Pioneer MTs and their plus-end tracking proteins (+TIPs) are thought to play integrative roles during this navigation. TACC3 is a + TIP that we have previously implicated in regulating MT dynamics within axons. However, the role of TACC3 in axon guidance has not been previously explored...
February 15, 2017: Neural Development
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