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https://www.readbyqxmd.com/read/29165592/folate-deficiency-induces-mitotic-aberrations-and-chromosomal-instability-by-compromising-the-spindle-assembly-checkpoint-in-cultured-human-colon-cells
#1
Xihan Guo, Juan Ni, Yuqian Zhu, Tao Zhou, Xiaoling Ma, Jinglun Xue, Xu Wang
Folates comprise the essential B9 vitamin that act as cofactors and cosubstrates in one-carbon metabolism for both biosynthesis and methylation of DNA and RNA. Folate deficiency (FD) has been shown to induce chromosomal instability (CIN), yet the underlying mechanisms are poorly understood. Here, we used human NCM460 colon mucosal cells as a model to investigate the effect of FD on spindle assembly checkpoint (SAC), a cell-cycle regulatory pathway preventing CIN during mitosis. Cells were maintained in medium containing 1...
November 19, 2017: Mutagenesis
https://www.readbyqxmd.com/read/29136577/atomic-force-microscopy-study-revealed-velocity-dependence-and-nonlinearity-of-nanoscale-poroelasticity-of-eukaryotic-cells
#2
Keyvan Mollaeian, Yi Liu, Siyu Bi, Juan Ren
Intracellular network deformation of the cell plays an important role in cellular shape formation. Recent studies suggest that cell reshaping and deformation due to external forces involve cellular volume, pore size, elasticity, and intracellular filaments polymerization degree change. This cell behavior can be described by poroelastic models due to the porous structure of the cytoplasm. In this study, the nanoscale poroelasticity of human mammary basal/claudin low carcinoma cell (MDA-MB-231) was investigated using indentation-based atomic force microscopy...
November 11, 2017: Journal of the Mechanical Behavior of Biomedical Materials
https://www.readbyqxmd.com/read/29134199/rapid-directed-transport-of-dc-sign-clusters-in-the-plasma-membrane
#3
Ping Liu, Violetta Weinreb, Marc Ridilla, Laurie Betts, Pratik Patel, Aravinda M de Silva, Nancy L Thompson, Ken Jacobson
C-type lectins, including dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN), are all-purpose pathogen receptors that exist in nanoclusters in plasma membranes of dendritic cells. A small fraction of these clusters, obvious from the videos, can undergo rapid, directed transport in the plane of the plasma membrane at average speeds of more than 1 μm/s in both dendritic cells and MX DC-SIGN murine fibroblasts ectopically expressing DC-SIGN. Surprisingly, instantaneous speeds can be considerably greater...
November 2017: Science Advances
https://www.readbyqxmd.com/read/29120369/begomoviral-movement-protein-effects-in-human-and-plant-cells-towards-new-potential-interaction-partners
#4
Susanna Krapp, Christian Schuy, Eva Greiner, Irina Stephan, Barbara Alberter, Christina Funk, Manfred Marschall, Christina Wege, Susanne M Bailer, Tatjana Kleinow, Björn Krenz
Geminiviral single-stranded circular DNA genomes replicate in nuclei so that the progeny DNA has to cross both the nuclear envelope and the plasmodesmata for systemic spread within plant tissues. For intra- and intercellular transport, two proteins are required: a nuclear shuttle protein (NSP) and a movement protein (MP). New characteristics of ectopically produced Abutilon mosaic virus (AbMV) MP (MP(AbMV)), either authentically expressed or fused to a yellow fluorescent protein or epitope tags, respectively, were determined by localization studies in mammalian cell lines in comparison to plant cells...
November 9, 2017: Viruses
https://www.readbyqxmd.com/read/29099266/calpain2-mediates-rab5-driven-focal-adhesion-disassembly-and-cell-migration
#5
Pablo A Mendoza, Patricio Silva, Jorge Díaz, Cecilia Arriagada, Jimena Canales, Oscar Cerda, Vicente A Torres
The early endosome protein Rab5 was recently shown to promote cell migration by enhancing focal adhesion disassembly through mechanisms that remain elusive. Focal adhesion disassembly is associated to proteolysis of talin, in a process that requires calpain2. Since calpain2 has been found at vesicles and endosomal compartments, we hypothesized that Rab5 stimulates calpain2 activity, leading to enhanced focal adhesion disassembly in migrating cells. We observed that calpain2 co-localizes with EEA1-positive early endosomes and co-immunoprecipitate with EEA1 and Rab5 in A549 lung carcinoma cells undergoing spreading, whereas Rab5 knock-down decreased the accumulation of calpain2 at early endosomal-enriched fractions...
November 3, 2017: Cell Adhesion & Migration
https://www.readbyqxmd.com/read/29092800/in-vivo-imaging-of-mauthner-axon-regeneration-remyelination-and-synapses-re-establishment-after-laser-axotomy-in-zebrafish-larvae
#6
Bing-Bing Hu, Min Chen, Rong-Chen Huang, Yu-Bin Huang, Yang Xu, Wu Yin, Lei Li, Bing Hu
Zebrafish is an excellent model to study central nervous system (CNS) axonal degeneration and regeneration since we can observe these processes in vivo and in real time in transparent larvae. Previous studies have shown that Mauthner cell (M-cell) axon regenerates poorly after mechanical spinal cord injury. Inconsistent with this result, however, we have found that M-cell possesses a great capacity for axon regeneration after two-photon laser ablation. By using ZEISS LSM 710 two-photon microscope, we performed specific unilateral axotomy of GFP labeled M-cells in the Tol-056 enhancer trap line larvae...
October 29, 2017: Experimental Neurology
https://www.readbyqxmd.com/read/29059502/effects-of-the-protonation-state-of-titratable-residues-and-the-presence-of-water-molecules-on-the-nocodazole-binding-to-%C3%AE-tubulin
#7
Dulce Consuelo Guzmán-Ocampo, Rodrigo Aguayo-Ortiz, Lucia Cano-González, Alicia Hernández-Campos, Rafael Castillo, Laura Dominguez
Regulation of microtubule assembly by antimitotic agents is a potential therapeutic strategy for the treatment of cancer, parasite infections, and neurodegenerative diseases. One of these agents is Nocodazole (NZ), which inhibits microtubule polymerization by binding to β-tubulin. NZ was recently co-crystallized in Gallus gallus tubulin, displaying novel information of the interaction features for ligand recognition and stability. In this work, we employed state of the art computational approaches to evaluate the protonation effects of titratable residues and the presence of water molecules in the binding of NZ...
October 23, 2017: ChemMedChem
https://www.readbyqxmd.com/read/29025057/nqo2-inhibition-relieves-reactive-oxygen-species-effects-on-mouse-oocyte-meiotic-maturation-and-embryo-development
#8
Dandan Chen, Xin Li, Xiaoyun Liu, Xiaoyu Liu, Xiuying Jiang, Juan Du, Qian Wang, Yuanjing Liang, Wei Ma
NRH: quinone oxidoreductase 2 (NQO2) is a cytosolic and ubiquitously expressed flavoprotein that catalyzes the two-electron reduction of quinone to hydroquinones. Herein, we assessed the protein expression, subcellular localization, and possible functions of NQO2 in mouse oocyte meiotic maturation and embryo development. Western blot analysis detected high and stable protein expression of NQO2 in mouse oocytes during meiotic progression. Immunofluorescence illustrated NQO2 distribution on nuclear membrane, chromosomes, and meiotic spindles...
October 1, 2017: Biology of Reproduction
https://www.readbyqxmd.com/read/29024608/erastin-like-anti-warburg-agents-prevent-mitochondrial-depolarization-induced-by-free-tubulin-and-decrease-lactate-formation-in-cancer-cells
#9
David N DeHart, John J Lemasters, Eduardo N Maldonado
In Warburg metabolism, suppression of mitochondrial metabolism contributes to a low cytosolic ATP/ADP ratio favoring enhanced aerobic glycolysis. Flux of metabolites across the mitochondrial outer membrane occurs through voltage-dependent anion channels (VDAC). In cancer cells, free dimeric tubulin induces VDAC closure and dynamically regulates mitochondrial membrane potential (ΔΨ). Erastin, a small molecule that binds to VDAC, antagonizes the inhibitory effect of tubulin on VDAC and hyperpolarizes mitochondria in intact cells...
September 1, 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28994750/use-of-immunolabeling-to-analyze-stable-dynamic-and-nascent-microtubules-in-the-zebrafish-embryo
#10
Rebecca J McFarland, Sharlene P Brown, Eudorah Vital, Jonathan M Werner, Rachel M Brewster
Microtubules (MTs) are dynamic and fragile structures that are challenging to image in vivo, particularly in vertebrate embryos. Immunolabeling methods are described here to analyze distinct populations of MTs in the developing neural tube of the zebrafish embryo. While the focus is on neural tissue, this methodology is broadly applicable to other tissues. The procedures are optimized for early to mid-somitogenesis-stage embryos (1 somite to 12 somites), however they can be adapted to a range of other stages with relatively minor adjustments...
September 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28985013/hdac2-3-binding-and-deacetylation-of-bubr1-initiates-the-spindle-assembly-checkpoint-silencing
#11
Inai Park, Mi-Sun Kwon, Sangjin Paik, Hyeonjong Kim, Hae-Ock Lee, Eunhee Choi, Hyunsook Lee
BubR1 acetylation is essential in spindle assembly checkpoint (SAC) signaling. Here we show that BubR1 deacetylation is a signal that initiates the mitotic exit. Sustained BubR1 acetylation arrests the cells in metaphase, although chromosome congression is achieved. BubR1 deacetylation was coordinated with dephosphorylation in mitotic exit, suggesting the presence of a coordinated acetylation-phosphorylation code in mitotic signaling. HDAC2 and 3 bound to acetylated BubR1 exclusively in mitosis and led to the polyubiquitination of BubR1...
October 6, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28950916/%C3%AE-carboxymuconolactone-decarboxylase-a-novel-cell-cycle-related-basal-body-protein-in-the-early-branching-eukaryote-trichomonas-vaginalis
#12
Wei-Hung Cheng, Kuo-Yang Huang, Po-Jung Huang, Chi-Ching Lee, Yuan-Ming Yeh, Fu-Man Ku, Rose Lin, Mei-Ling Cheng, Cheng-Hsun Chiu, Petrus Tang
BACKGROUND: γ-Carboxymuconolactone decarboxylase (CMD) participates in the β-ketoadipate pathway, which catalyzes aromatic compounds to produce acetyl- or succinyl-CoA, in prokaryotes and yeast. Our previous study demonstrated that expression of a CMD homologue that contains two signatures (dualCMD) is negatively regulated by iron in Trichomonas vaginalis. However, we were not able to identify the components of the β-ketoadipate pathway in the parasite's genome. These observations prompted us to investigate the biological functions of this novel CMD homologue in T...
September 26, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28942021/a-novel-immunofluorescence-method-to-visualize-microtubules-in-the-antiparallel-overlaps-of-microtubule-plus-ends-in-the-anaphase-and-telophase-midzone
#13
Aya Ifuji, Takahisa Kuga, Yuichiro Kaibori, Youhei Saito, Yuji Nakayama
Cell division, in which duplicated chromosomes are separated into two daughter cells, is the most dynamic event during cell proliferation. Chromosome movement is powered mainly by microtubules, which vary in morphology and are organized into characteristic structures according to mitotic progression. During the later stages of mitosis, antiparallel microtubules form the spindle midzone, and the irregular formation of the midzone often leads to failure of cytokinesis, giving rise to the unequal segregation of chromosomes...
November 15, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28934466/cohesin-acetyltransferase-esco2-regulates-sac-and-kinetochore-functions-via-maintaining-h4k16-acetylation-during-mouse-oocyte-meiosis
#14
Yajuan Lu, Xiaoxin Dai, Mianqun Zhang, Yilong Miao, Changyin Zhou, Zhaokang Cui, Bo Xiong
Sister chromatid cohesion, mediated by cohesin complex and established by the acetyltransferases Esco1 and Esco2, is essential for faithful chromosome segregation. Mutations in Esco2 cause Roberts syndrome, a developmental disease characterized by severe prenatal retardation as well as limb and facial abnormalities. However, its exact roles during oocyte meiosis have not clearly defined. Here, we report that Esco2 localizes to the chromosomes during oocyte meiotic maturation. Depletion of Esco2 by morpholino microinjection leads to the precocious polar body extrusion, the escape of metaphase I arrest induced by nocodazole treatment and the loss of BubR1 from kinetochores, indicative of inactivated SAC...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28928489/the-phosphorylation-of-a-kinetochore-protein-dam1-by-aurora-b-ipl1-kinase-promotes-chromosome-bipolar-attachment-in-yeast
#15
Fengzhi Jin, Michael Bokros, Yanchang Wang
The interaction between chromosomes and spindle microtubules is essential for chromosome segregation. The kinetochore complex mediates this interaction. Previous studies indicate that the stability of kinetochore attachment is regulated by Aurora B/Ipl1 kinase and this regulation is conserved from yeast to mammalian cells. In budding yeast Saccharomyces cerevisiae, the ten-subunit Dam1/DASH complex bridges the interaction between kinetochores and microtubules, and some in vitro evidence indicates that the phosphorylation of Dam1 protein by Ipl1 kinase destabilizes this interaction...
September 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28916777/the-catalytic-subunit-of-dna-polymerase-%C3%AE-inhibits-%C3%AE-turc-activity-and-regulates-golgi-derived-microtubules
#16
Yuehong Shen, Pengfei Liu, Taolue Jiang, Yu Hu, Franco K C Au, Robert Z Qi
γ-Tubulin ring complexes (γTuRCs) initiate microtubule growth and mediate microtubule attachment at microtubule-organizing centers, such as centrosomes and the Golgi complex. However, the mechanisms that control γTuRC-mediated microtubule nucleation have remained mostly unknown. Here, we show that the DNA polymerase δ catalytic subunit (PolD1) binds directly to γTuRCs and potently inhibits γTuRC-mediated microtubule nucleation. Whereas PolD1 depletion through RNA interference does not influence centrosome-based microtubule growth, the depletion augments microtubule nucleation at the Golgi complex...
September 15, 2017: Nature Communications
https://www.readbyqxmd.com/read/28889243/a-high-throughput-qcm-chip-configuration-for-the-study-of-living-cells-and-cell-drug-interactions
#17
Haibo Shen, Tiean Zhou, Jiajin Hu
In this study, we present a novel design of interference-free, negligible installation-induced stress, suitable for the fabrication of high-throughput quartz crystal microbalance (HQCM) chips. This novel HQCM chip configuration was fabricated using eight independent yet same-batch quartz crystal resonators within a common glass substrate with eight through-holes of diameter slightly larger than that of the quartz resonator. Each quartz resonator's rim was adhered to the inner part of the through-hole via silicone glue to form the rigid (quartz)-soft (silicone)-rigid (glass) structure (RSRS) which effectively eliminates the acoustic couplings among different resonators and largely alleviates the installation-induced stresses...
September 9, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28884019/microtubule-binding-protein-for20-promotes-microtubule-depolymerization-and-cell-migration
#18
Sijie Feng, Yinlong Song, Minhong Shen, Shanshan Xie, Wenjing Li, Yi Lu, Yuehong Yang, Guangshuo Ou, Jun Zhou, Fudi Wang, Wei Liu, Xiaoyi Yan, Xin Liang, Tianhua Zhou
Microtubules are highly dynamic filaments assembled from αβ-tubulin heterodimers and play important roles in many cellular processes, including cell division and migration. Microtubule dynamics is tightly regulated by microtubule-associated proteins (MAPs) that function by binding to microtubules or free tubulin dimers. Here, we report that FOR20 (FOP-related protein of 20 kDa), a conserved protein critical for ciliogenesis and cell cycle progression, is a previously uncharacterized MAP that facilitates microtubule depolymerization and promotes cell migration...
2017: Cell Discovery
https://www.readbyqxmd.com/read/28878350/functional-association-of-cellular-microtubules-with-viral-capsid-assembly-supports-efficient-hepatitis-b-virus-replication
#19
Masashi Iwamoto, Dawei Cai, Masaya Sugiyama, Ryosuke Suzuki, Hideki Aizaki, Akihide Ryo, Naoko Ohtani, Yasuhito Tanaka, Masashi Mizokami, Takaji Wakita, Haitao Guo, Koichi Watashi
Viruses exploit host factors and environment for their efficient replication. The virus-host interaction mechanisms for achieving an optimal hepatitis B virus (HBV) replication have been largely unknown. Here, a single cell cloning revealed that HepAD38 cells, a widely-used HBV-inducible cell line, contain cell clones with diverse permissiveness to HBV replication. The HBV permissiveness was impaired upon treatment with microtubule inhibitor nocodazole, which was identified as an HBV replication inhibitor from a pharmacological screening...
September 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28855121/tubulin-is-retained-throughout-the-human-hematopoietic-erythroid-cell-differentiation-process-and-plays-a-structural-role-in-sedimentable-fraction-of-mature-erythrocytes
#20
Ayelén D Nigra, Verónica S Santander, Roberto Dircio-Maldonado, Marina Rafaela Amaiden, Noelia E Monesterolo, Patricia Flores-Guzmán, Tamara Muhlberger, Juan F Rivelli, Alexis N Campetelli, Héctor Mayani, Cesar H Casale
We investigated the properties of tubulin present in the sedimentable fraction ("Sed-tub") of human erythrocytes, and tracked the location and organization of tubulin in various types of cells during the process of hematopoietic/erythroid differentiation. Sed-tub was sensitive to taxol/nocodazole (drugs that modify microtubule assembly/disassembly), but was organized as part of a protein network rather than in typical microtubule form. This network had a non-uniform "connected-ring" structure, with tubulin localized in the connection areas and associated with other proteins...
October 2017: International Journal of Biochemistry & Cell Biology
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