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Cytoskeleton

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https://www.readbyqxmd.com/read/28407416/tau-based-fluorescent-protein-fusions-to-visualize-microtubules
#1
Paul Mooney, Taylor Sulerud, James Pelletier, Matthew Dilsaver, Miroslav Tomschik, Christoph Geisler, Jesse C Gatlin
The ability to visualize cytoskeletal proteins and their dynamics in living cells has been critically important in advancing our understanding of numerous cellular processes, including actin- and microtubule-dependent phenomena such as cell motility, cell division, and mitosis. Here we describe a novel set of fluorescent protein fusions designed specifically to visualize microtubules in living systems using fluorescence microscopy. Each fusion contains a fluorescent protein module linked in frame to a modified phospho-deficient version of the microtubule-binding domain of Tau (mTMBD)...
April 13, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28378936/the-tumor-suppressor-tropomyosin-tpm2-1-regulates-sensitivity-to-apoptosis-beyond-anoikis-characterized-by-changes-in-the-levels-of-intrinsic-apoptosis-proteins
#2
Melissa Desouza-Armstrong, Peter W Gunning, Justine R Stehn
The actin cytoskeleton is a polymer system that acts both as a sensor and mediator of apoptosis. Tropomyosins (Tpm) are a family of actin binding proteins that form co-polymers with actin and diversify actin filament function. Previous studies have shown that elevated expression of the tropomyosin isoform Tpm2.1 sensitized cells to apoptosis induced by cell detachment (anoikis) via an unknown mechanism. It is not yet known whether Tpm2.1 or other tropomyosin isoforms regulate sensitivity to apoptosis beyond anoikis...
April 5, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28329908/re-evaluating-the-roles-of-myosin-18a%C3%AE-and-f-actin-in-determining-golgi-morphology
#3
Kyle Bruun, Jordan R Beach, Sarah M Heissler, Kirsten Remmert, James R Sellers, John A Hammer
The peri-centrosomal localization and morphology of the Golgi apparatus depends largely on the microtubule cytoskeleton and the microtubule motor protein dynein. Recent studies proposed that myosin 18Aα (M18Aα) also contributes to Golgi morphology by binding the Golgi protein GOLPH3 and walking along adjacent actin filaments to stretch the Golgi into its classic ribbon structure. Biochemical analyses have shown, however, that M18A is not an actin-activated ATPase and lacks motor activity. Our goal, therefore, was to define the precise molecular mechanism by which M18Aα determines Golgi morphology...
March 22, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28317275/synthesis-of-%C3%AE-tubulin-occurs-within-chromatoid-body-of-round-spermatids
#4
Yuki Fujii, Hideaki Fujita, Sadaki Yokota
mRNAs for proteins required in elongated spermatids are considered to be transcribed at an early stage and stored in cytoplasm, presumably in chromatoid body (CB), one type of nuage component (a unique structure that appears and disappears during spermatogenesis), because transcription of genes does not occur at late stages. In elongated spermatids, a large amount of tubulin molecules is required to form microtubules of manchette and flagellum. To investigate the possible role of CB in translation of tubulin mRNA, we performed immunofluorescence and immunoelectron microscopic localization studies of α- and β-tubulin in rat spermatogenic cells...
March 20, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28296067/rna-binding-proteins-of-the-nxf-nuclear-export-factor-family-and-their-connection-with-the-cytoskeleton
#5
REVIEW
L A Mamon, V R Ginanova, S F Kliver, A O Yakimova, A A Atsapkina, E V Golubkova
The mutual relationship between mRNA and the cytoskeleton can be seen from two points of view. On the one hand, the cytoskeleton is necessary for mRNA trafficking and anchoring to subcellular domains. On the other hand, cytoskeletal growth and rearrangement require the translation of mRNAs that are connected to the cytoskeleton. β-actin mRNA localization may influence dynamic changes in the actin cytoskeleton. In the cytoplasm, long-lived mRNAs exist in the form of RNP (ribonucleoprotein) complexes, where they interact with RNA-binding proteins, including NXF (Nuclear eXport Factor)...
March 13, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28236372/the-rotation-of-mouse-myoblast-nuclei-is-dependent-on-substrate-elasticity
#6
Ryan Hickey, Andrew E Pelling
The complex interplay of biochemical signaling and mechanical traction forces regulate the position of cellular nuclei. Although the phenomenon of nuclear rotation has been observed for many years, the influence of substrate elasticity was unknown. We discovered another layer of complexity to this phenomenon: nuclear rotation is dependent on substrate elasticity. Nuclear rotation is drastically reduced on physiologically relevant stiffnesses. Here, we studied nuclear rotation in mouse C2C12 myoblasts cultured on soft substrates designed to mimic resting tissue (∼26 kPa) and on hard glass substrates...
February 25, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28218453/divergent-roles-of-%C3%AE-and-%C3%AE-actin-isoforms-during-spread-of-vaccinia-virus
#7
N Bishara Marzook, Sharissa L Latham, Helena Lynn, Christopher Mckenzie, Christine Chaponnier, Georges E Grau, Timothy P Newsome
Actin is a major component of the cytoskeleton and is present as two isoforms in non-muscle cells: β- and γ-cytoplasmic actin. These isoforms are strikingly conserved, differing by only four N-terminal amino acids. During spread from infected cells, vaccinia virus (VACV) particles induce localized actin nucleation that propel virus to surrounding cells and facilitate cell-to-cell spread of infection. Here we show that virus-tipped actin comets are composed of β- and γ-actin. We employed isoform-specific siRNA knockdown to examine the role of the two isoforms in VACV-induced actin comets...
February 20, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28164467/microtubule-regulating-proteins-and-camp-dependent-signaling-in-neuroblastoma-differentiation
#8
Pablo Muñoz-Llancao, Cristian de Gregorio, Macarena Las Heras, Christopher Meinohl, Kevin Noorman, Erik Boddeke, Xiaodong Cheng, Frank Lezoualc'h, Martina Schmidt, Christian Gonzalez-Billault
Neurons are highly differentiated cells responsible for the conduction and transmission of information in the nervous system. The proper function of a neuron relies on the compartmentalization of their intracellular domains. Differentiated neuroblastoma cells have been extensively used to study and understand the physiology and cell biology of neuronal cells. Here, we show that differentiation of N1E-115 neuroblastoma cells is more pronounced upon exposure of a chemical analog of cyclic AMP (cAMP), db-cAMP...
March 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28063204/assembly-studies-of-six-intestinal-intermediate-filament-if-proteins-b2-c1-c2-d1-d2-and-e1-in-the-nematode-c-elegans
#9
Anton Karabinos, Jürgen Schünemann, David A D Parry
The dimerisation properties of six intestine-expressed intermediate filament (IF) proteins (B2, C1, C2, D1, D2, E1) were analysed in blot overlay assay on membranes containing all of the eleven recombinant C. elegans IF proteins (A1, A2, A3, A4, B1, B2, C1, C2, D1, D2, and E1). The interactions detected in the blot assays exclusively comprise intestine-expressed IF proteins and the protein A4, which is found in the dauer larva intestine. About 86% of these interactions are heterotypic, while the remaining interactions relate to C1, C2, and D2 homodimers...
March 2017: Cytoskeleton
https://www.readbyqxmd.com/read/27998020/identification-characterization-and-expression-of-sarcomeric-tropomyosin-isoforms-in-zebrafish
#10
Dipak K Dube, Syamalima Dube, Lynn Abbott, Jushuo Wang, Yingli Fan, Ruham Alshiekh-Nasany, Kalpesh K Shah, Alexander P Rudloff, Bernard J Poiesz, Jean M Sanger, Joseph W Sanger
Tropomyosin is a component of thin filaments that constitute myofibrils, the contractile apparatus of striated muscles. In vertebrates, except for fish, four TPM genes TPM1, TPM2, TPM3, and TPM4 are known. In zebrafish, there are six TPM genes that include the paralogs of the TPM1 (TPM1-1 and TPM1-2), the paralogs of the TPM4 gene (TPM4-1 and TPM4-2), and the two single copy genes TPM2 and TPM3. In this study, we have identified, cloned, and sequenced the TPM1-1κ isoform of the TPM1-1 gene and also discovered a new isoform TPM1-2ν of the TPM1-2...
March 2017: Cytoskeleton
https://www.readbyqxmd.com/read/27935261/matrix-rigidity-regulates-microtubule-network-polarization-in-migration
#11
Matthew Raab, Dennis E Discher
The microtubule organizing center (MTOC) frequently polarizes to a position in front of the nucleus during cell migration, but recent work has shown conflicting evidence for MTOC location in migratory polarized cells. Here, we show that subcellular localization of the MTOC is modulated by extracellular matrix stiffness. In scratch wound assays as well as single cell migration of mesenchymal stem cells (MSCs) the MTOC appears randomly positioned when cells are migrating on soft matrix, whereas on stiff matrix the MTOC is in front of the nucleus...
March 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28026138/towards-a-molecular-architecture-of-the-centrosome-in-toxoplasma-gondii
#12
REVIEW
Juliette Morlon-Guyot, Maria E Francia, Jean-François Dubremetz, Wassim Daher
Toxoplasma gondii is the causative agent of toxoplasmosis. The pathogenicity of this unicellular parasite is tightly linked to its ability to efficiently proliferate within its host. Tachyzoites, the fast dividing form of the parasite, divide by endodyogeny. This process involves a single round of DNA replication, closed nuclear mitosis, and assembly of two daughter cells within a mother. The successful completion of endodyogeny relies on the temporal and spatial coordination of a plethora of simultaneous events...
February 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28001338/wdpcp-regulates-the-ciliogenesis-of-human-sinonasal-epithelial-cells-in-chronic-rhinosinusitis
#13
Yun Ma, Yueqi Sun, Lijie Jiang, Kejun Zuo, Hexin Chen, Jiebo Guo, Fenghong Chen, Yinyan Lai, Jianbo Shi
Damage to the mucociliary clearance system is a typical change in the pathogenesis in chronic rhinosinusitis. However, the mechanisms underlying cilia loss remain unclear. WDPCP is a key protein essential for ciliogenesis, and is also an effector of the planar cell polarity signaling system. In this study, we sought to determine the role of WDPCP in cilia loss in patients with chronic rhinosinusitis. We demonstrated the expression of WDPCP in human sinonasal epithelium from patients with chronic rhinosinusitis and control subjects...
February 2017: Cytoskeleton
https://www.readbyqxmd.com/read/27997747/dynamics-of-the-sealing-zone-in-cultured-osteoclasts
#14
Sarit Batsir, Benjamin Geiger, Zvi Kam
Bone resorption by osteoclasts (OCs) depends on the formation and stability of the sealing zone (SZ), a peripheral belt of actin and integrin-based podosomes. Recent studies demonstrated that the SZ is a highly dynamic structure, undergoing cycles of assembly and disassembly. In this study, we explored the mechanisms underlying the regulation of SZ stability and reorganization in OCs cultured on glass slides, and forming an SZ-like podosome belt (SZL). By monitoring this belt in cultured RAW264.7 cells expressing GFP-tagged actin, we show here that SZL stability is usually locally regulated, and its dissociation, occurring mostly in concave segments, is manifested in the loss of both podosome coherence, and actin belt continuity...
February 2017: Cytoskeleton
https://www.readbyqxmd.com/read/27935262/elastic-tethers-between-separating-anaphase-chromosomes-in-crane-fly-spermatocytes-coordinate-chromosome-movements-to-the-two-poles
#15
Rozhan Sheykhani, Michael Berns, Arthur Forer
Separating anaphase chromosomes in crane-fly spermatocytes are connected by elastic tethers, as originally described by LaFountain et al. (2002): telomere-containing arm fragments severed from the arms move backwards to the partner telomeres. We have tested whether the tethers coordinate the movements of separating partner chromosomes. In other cell types anaphase chromosomes move faster, temporarily, when their kinetochore microtubules are severed. However, in crane-fly spermatocytes the chromosomes move at their usual speed when their kinetochore microtubules are severed...
February 2017: Cytoskeleton
https://www.readbyqxmd.com/read/27935245/modeling-the-effects-of-lattice-defects-on-microtubule-breaking-and-healing
#16
Nan Jiang, Megan E Bailey, Jessica Burke, Jennifer L Ross, Ruxandra I Dima
Microtubule reorganization often results from the loss of polymer induced through break- age or active destruction by energy-using enzymes. Pre-existing defects in the microtubule lattice likely lower structural integrity and aid filament destruction. Using large-scale molecular simulations, we model diverse microtubule fragments under forces generated at specific positions to locally crush the filament. We show that lattices with 2% defects are crushed and severed by forces three times smaller than defect-free ones...
December 9, 2016: Cytoskeleton
https://www.readbyqxmd.com/read/27935233/kinetic-analysis-of-de-novo-centriole-assembly-in-heat-shocked-mammalian-cells
#17
In Keol Baek, Yeun Kyu Jang, Tae H Lee, JooHun Lee
Mammalian cells are capable of de novo centriole formation after the removal of existing centrioles. This suggests that de novo centriole assembly is repressed in normally duplicating cells to maintain a constant number of centrioles in the cells. However, neither the mechanism of de novo centriole assembly nor that of its hypothesized repression is understood due to the lack of an experimental system. We found that the heat shock (HS; 42°C, 2 h) of mouse embryonic fibroblasts caused the separation of centriole pairs, a transient increase in polo-like kinase (Plk) 4 expression, and the formation of a complex containing γ-tubulin, pericentrin, HS protein (Hsp) 90, and Plk4, in approximately half of the cells...
December 9, 2016: Cytoskeleton
https://www.readbyqxmd.com/read/27783462/effect-of-surface-chemistry-on-tropomyosin-binding-to-actin-filaments-on-surfaces
#18
Philip R Nicovich, Miro Janco, Tom Sobey, Mehul Gajwani, Peyman Obeidy, Renee Whan, Katharina Gaus, Peter W Gunning, Adelle Cf Coster, Till Böcking
Reconstitution of actin filaments on surfaces for observation of filament-associated protein dynamics by fluorescence microscopy is currently an exciting field in biophysics. Here we examine the effects of attaching actin filaments to surfaces on the binding and dissociation kinetics of a fluorescence-labeled tropomyosin, a rod-shaped protein that forms continuous strands wrapping around the actin filament. Two attachment modalities of the actin to the surface are explored: where the actin filament is attached to the surface at multiple points along its length; and where the actin filament is attached at one end and aligned parallel to the surface by buffer flow...
December 2016: Cytoskeleton
https://www.readbyqxmd.com/read/27770600/inf2-and-fhod-related-formins-promote-ovulation-in-the-somatic-gonad-of-c-elegans
#19
Anna Hegsted, Forrest A Wright, SarahBeth Votra, David Pruyne
Formins are regulators of actin filament dynamics. We demonstrate here that two formins, FHOD-1 and EXC-6, are important in the nematode Caenorhabditis elegans for ovulation, during which actomyosin contractions push a maturing oocyte from the gonad arm into a distensible bag-like organ, the spermatheca. EXC-6, a homolog of the disease-associated mammalian formin INF2, is highly expressed in the spermatheca, where it localizes to cell-cell junctions and to circumferential actin filament bundles. Loss of EXC-6 does not noticeably affect the organization the actin filament bundles, and causes only a very modest increase in the population of junction-associated actin filaments...
December 2016: Cytoskeleton
https://www.readbyqxmd.com/read/27770595/microtubule-binding-protein-pacrg-plays-a-role-in-regulating-specific-ciliary-dyneins-during-microtubule-sliding
#20
Katsutoshi Mizuno, Erin E Dymek, Elizabeth F Smith
The complex waveforms characteristic of motile eukaryotic cilia and flagella are produced by the temporally and spatially regulated action of multiple dynein subforms generating sliding between subsets of axonemal microtubules. Multiple protein complexes have been identified that are associated with the doublet microtubules and that mediate regulatory signals between key axonemal structures, such as the radial spokes and central apparatus, and the dynein arm motors; these complexes include the N-DRC, MIA, and CSC complexes...
December 2016: Cytoskeleton
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