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https://www.readbyqxmd.com/read/28722126/investigating-the-effect-of-poly-l-lactic-acid-nanoparticles-carrying-hypericin-on-the-flow-biased-diffusive-motion-of-hela-cell-organelles
#1
Rozhin Penjweini, Sarah Deville, Omid Haji Maghsoudi, Kristof Notelaers, Anitha Ethirajan, Marcel Ameloot
OBJECTIVES: In this study, we investigate in human cervical epithelial HeLa cells the intracellular dynamics and the mutual interaction with the organelles of the poly-l-lactic acid nanoparticles (PLLA NPs) carrying the naturally occurring hydrophobic photosensitizer hypericin. METHODS: Temporal and spatiotemporal image correlation spectroscopy was used for the assessment of the intracellular diffusion and directed motion of the nanocarriers by tracking the hypericin fluorescence...
July 19, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/28720756/pixuna-virus-modifies-host-cell-cytoskeleton-to-secure-infection
#2
Pedro Ignacio Gil, Guillermo Albrieu-Llinás, Estela Cecilia Mlewski, Marina Monetti, Laura Fozzatti, Cecilia Cuffini, José Fernández Romero, Patricia Kunda, María Gabriela Paglini
Pixuna virus (PIXV) is an enzootic member of the Venezuelan Equine Encephalitis Virus complex and belongs to the New World cluster of alphaviruses. Herein we explore the role of the cellular cytoskeleton during PIXV replication. We first identified that PIXV undergoes an eclipse phase consisting of 4 h followed by 20 h of an exponential phase in Vero cells. The infected cells showed morphological changes due to structural modifications in actin microfilaments (MFs) and microtubules (MTs). Cytoskeleton-binding agents, that alter the architecture and dynamics of MFs and MTs, were used to study the role of cytoskeleton on PIXV replication...
July 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28701724/dynamic-microtubule-association-of-doublecortin-x-dcx-is-regulated-by-its-c-terminus
#3
Maryam Moslehi, Dominic C H Ng, Marie A Bogoyevitch
Doublecortin X (DCX), known to be essential for neuronal migration and cortical layering in the developing brain, is a 40 kDa microtubule (MT)-associated protein. DCX directly interacts with MTs via its two structured doublecortin (DC) domains, but the dynamics of this association and the possible regulatory roles played by the flanking unstructured regions remain poorly defined. Here, we employ quantitative fluorescence recovery after photobleaching (FRAP) protocols in living cells to reveal that DCX shows remarkably rapid and complete exchange within the MT network but that the removal of the C-terminal region significantly slows this exchange...
July 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28694353/a-centrosomal-protein-for20-regulates-microtubule-assembly-dynamics-and-plays-a-role-in-cell-migration
#4
Shalini Srivastava, Dulal Panda
Here, we report that a centrosomal protein FOR20 (also named as C16orf63, FLJ31153 or PHSECRG2) can regulate the assembly and stability of microtubules. Both FOR20 IgG antibody and GST-tagged FOR20 could precipitate tubulin from HeLa cell extract indicating a possible interaction between FOR20 and tubulin. FOR20 was also detected in goat brain tissue extract and it cycled with microtubule-associated proteins. Further, FOR20 bound to purified tubulin and inhibited the assembly of tubulin in vitro. The over-expression of FOR20 depolymerized interphase microtubules and the depletion of FOR20 prevented nocodazole-induced depolymerization of microtubules in HeLa cells...
July 10, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28684120/design-and-synthesis-of-4-morpholino-6-1-2-3-6-tetrahydropyridin-4-yl-n-3-4-5-trimethoxyphenyl-1-3-5-triazin-2-amine-analogues-as-tubulin-polymerization-inhibitors
#5
Suresh Narva, Surendar Chitti, Suresh Amaroju, Debanjan Bhattacharjee, Bala Bhaskara Rao, Nishant Jain, Mallika Alvala, Kondapalli Venkata Gowri Chandra Sekhar
A series of thirty-seven 1,3,5-triazine analogues have been synthesized, characterized and evaluated for their antiproliferative activity against a panel of four different human cancer cell lines such as HeLa, HepG2, A549 and MCF-7. Most of the 1,3,5-triazine analogues exhibited promising antiproliferative activity against tested cancer cell lines. Among all the synthesized compounds, 8j showed potent activity against the cancer cell lines such as HeLa, HepG2, A549 and MCF-7 with IC50 12.3±0.8, 9.6±0.4, 10...
June 24, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28654080/studying-cell-cycle-regulated-gene-expression-by-two-complementary-cell-synchronization-protocols
#6
Aintzane Apraiz, Jone Mitxelena, Ana Zubiaga
The gene expression program of the cell cycle represents a critical step for understanding cell cycle-dependent processes and their role in diseases such as cancer. Cell cycle-regulated gene expression analysis depends on cell synchronization into specific phases. Here we describe a method utilizing two complementary synchronization protocols that is commonly used for studying periodic variation of gene expression during the cell cycle. Both procedures are based on transiently blocking the cell cycle in one defined point...
June 6, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28651607/the-long-non-coding-rna-bc200-bcyrn1-is-critical-for-cancer-cell-survival-and-proliferation
#7
Evan P Booy, Ewan Ks McRae, Amit Koul, Francis Lin, Sean A McKenna
BACKGROUND: BC200 is a long non-coding RNA expressed at high levels in the brain and elevated in a variety of tumour types. BC200 has a hypothesized role in translational regulation; however, to date the functional role of BC200 in both normal and diseased states remains poorly characterized. METHODS: Detailed BC200 expression analyses were performed in tumor cell lines, primary and non-tumorigenic cultured breast and lung cells, and a panel of normal human tissues by quantitative real-time PCR and confirmed by northern blot...
June 26, 2017: Molecular Cancer
https://www.readbyqxmd.com/read/28643946/activated-proline-rich-tyrosine-kinase-2-regulates-meiotic-spindle-assembly-in-the-mouse-oocyte
#8
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
#9
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
#10
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
#11
Kohgaku Eguchi, Zacharie Taoufiq, Oliver Thorn-Seshold, Dirk Trauner, Masato Hasegawa, Tomoyuki Takahashi
α-Synuclein is a presynaptic protein the function of which has 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 sex...
June 21, 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
#12
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...
June 18, 2017: Cell Cycle
https://www.readbyqxmd.com/read/28557611/lamin-a-and-microtubules-collaborate-to-maintain-nuclear-morphology
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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...
August 2017: Histochemistry and Cell Biology
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