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Molecular Biology of the Cell

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https://www.readbyqxmd.com/read/28077622/ciliary-entry-of-kif17-is-dependent-on-its-binding-to-the-ift-b-complex-via-ift46-ift56-as-well-as-on-its-nuclear-localization-signal
#1
Teruki Funabashi, Yohei Katoh, Saki Michisaka, Masaya Terada, Maho Sugawa, Kazuhisa Nakayama
Cilia function as cellular antennae to sense and transduce extracellular signals. A number of proteins are specifically localized in cilia. Anterograde and retrograde ciliary protein trafficking are mediated by the IFT-B and IFT-A complexes in concert with kinesin-2 and dynein-2 motors, respectively. However, the role of KIF17, a homodimeric kinesin-2 protein, in protein trafficking has not been fully understood in vertebrate cilia. In this study, we first demonstrated by taking advantage of the visible immunoprecipitation assay, that KIF17 interacts with the IFT46-IFT56 dimer in the IFT-B complex through its C-terminal sequence located immediately upstream of the nuclear localization signal (NLS)...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077621/functional-redundancy-and-non-redundancy-between-two-troponin-c-isoforms-in-drosophila-adult-muscles
#2
Maria B Chechenova, Sara Maes, Sandy T Oas, Cloyce Nelson, Kaveh G Kiani, Anton L Bryantsev, Richard M Cripps
We investigated the functional overlap of two muscle Troponin C (TpnC) genes that are expressed in the adult fruit fly, Drosophila melanogaster: TpnC4 is predominantly expressed in the indirect flight muscles (IFM), whereas TpnC41C is the main isoform in the tergal depressor of the trochanter muscle (TDT or jump muscle). Using CRISPR/Cas9, we created a transgenic line with a homozygous deletion of TpnC41C, and compared its phenotype to a line lacking functional TpnC4 We found that the removal of either of these genes leads to expression of the other isoform in both muscle types...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077620/mitogen-induced-distinct-epi-alleles-are-phosphorylated-at-either-h3s10-or-h3s28-depending-on-h3k27-acetylation
#3
Dilshad H Khan, Shannon Healy, Shihua He, Daniel Lichtensztejn, Ludger Klewes, Kiran L Sharma, Veronica Lau, Sabine Mai, Geneviève P Delcuve, James R Davie
Stimulation of the MAPK pathway results in mitogen- and stress-activated protein kinase 1/2 (MSK1/2)-catalyzed phosphorylation of histone H3 at serine 10 or 28 and expression of immediate-early (IE) genes. In 10T1/2 mouse fibroblasts, phosphorylation of H3S10 and H3S28 occurs on different H3 molecules and in different nuclear regions. Likewise, we show that mitogen-induced H3S10 and H3S28 phosphorylation occurs in separate pools in human primary fibroblasts. High-resolution imaging studies on both cell types have revealed that H3S10 and H3S28 phosphorylation events can be induced in a single cell but on different alleles, giving rise to H3S10ph and H3S28ph epi-alleles...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077619/relative-importance-of-%C3%AE-cyto-and-%C3%AE-cyto-actin-in-primary-mouse-embryonic-fibroblasts
#4
Xiaobai Patrinostro, Allison R O'Rourke, Christopher M Chamberlain, Branden S Moriarity, Benjamin J Perrin, James M Ervasti
The highly homologous beta (βcyto) and gamma (γcyto) cytoplasmic actins are hypothesized to carry out both redundant and unique essential functions, but studies using targeted gene knockout and siRNA mediated transcript knockdown to examine βcyto- and γcyto-isoform specific functions in various cell types have yielded conflicting data. Here, we quantitatively characterized actin transcript and protein levels as well as cellular phenotypes in both gene- and transcript-targeted primary mouse embryonic fibroblasts...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077618/mechanism-of-nuclear-movements-in-a-multinucleated-cell
#5
Romain Gibeaux, Antonio Z Politi, Peter Philippsen, François Nédélec
Multinucleated cells are important in many organisms but the mechanisms governing the movements of nuclei sharing a common cytoplasm are not understood. In the hyphae of the plant pathogenic fungus Ashbya gossypii, nuclei move back and forth, occasionally bypassing each other, and, preventing the formation of nuclear clusters, this is essential for genetic stability. These movements depend on cytoplasmic microtubules emanating from the nuclei, that are pulled by dynein motors anchored at the cortex. Using 3D stochastic simulations with parameters constrained by the literature, we predict the cortical anchors density from the characteristics of nuclear movements...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077617/calcineurin-the-ca2-dependent-phosphatase-regulates-rga2-a-cdc42-gtpase-activating-protein-to-modulate-pheromone-signaling
#6
Nina Ly, Martha S Cyert
Calcineurin, the conserved Ca(2+)/calmodulin-activated phosphatase, is required for viability during prolonged exposure to pheromone, and acts through multiple substrates to down regulate yeast pheromone signaling. Calcineurin regulates Dig2 and Rod1/Art4 to inhibit mating-induced gene expression and activate receptor internalization, respectively. Recent systematic approaches identified Rga2, a GTPase activating protein (GAP) for the Cdc42 Rho-type GTPase, as a calcineurin substrate. Here, we establish a physiological context for this regulation, and show that calcineurin dephosphorylates and positively regulates Rga2 during pheromone signaling...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28077616/different-modes-of-enhancer-specific-regulation-by-runt-and-even-skipped-during-drosophila-segmentation
#7
Saiyu Hang, J Peter Gergen
The initial metameric expression of the Drosophila sloppy paired 1 (slp1) gene is controlled by two distinct cis-regulatory DNA elements that interact in a non-additive manner to integrate inputs from transcription factors encoded by the pair-rule segmentation genes. We performed Chromatin Immuno-Precipitation (ChIP) on reporter genes containing these elements in different embryonic genotypes to investigate the mechanism of their regulation. The Distal Early Stripe Element (DESE) mediates both activation and repression by Runt...
January 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057767/nano-random-forests-to-mine-protein-complexes-and-their-relationships-in-quantitative-proteomics-data
#8
Luis F Montaño-Gutierrez, Shinya Ohta, Georg Kustatscher, William C Earnshaw, Juri Rappsilber
Ever-increasing numbers of quantitative proteomics datasets constitute a currently underexploited resource for investigating protein function. Multi-protein complexes often follow consistent trends in these experiments, which could provide insights about their biology. Yet, as more experiments are considered, a complex's signature may become conditional and less identifiable. Previously, we successfully distinguished the general proteomic signature of genuine chromosomal proteins from hitchhikers using the Random Forests (RFs) machine learning algorithm...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057766/rapid-degradation-of-mutant-slc25a46-by-the-ubiquitin-proteasome-system-results-in-mfn1-2-mediated-hyperfusion-of-mitochondria
#9
Janos Steffen, Ajay A Vashisht, Jijun Wan, Joanna C Jen, Steven M Claypool, James A Wohlschlegel, Carla M Koehler
SCL25A46 is a member of the mitochondrial carrier protein that surprisingly localizes to the outer membrane and is distantly related to Ugo1. Here we show that a subset of SLC25A46 interacted with mitochondrial dynamics components and the MICOS complex. Decreased expression of SLC25A46 resulted in increased stability and oligomerization of MFN1 and MFN2 on mitochondria, promoting mitochondrial hyperfusion. A mutation at L341P caused rapid degradation of SLC25A46 that manifested as a rare disease, pontocerebellar hypoplasia...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057765/insulin-signaling-regulates-a-functional-interaction-between-adenomatous-polyposis-coli-apc-and-cytoplasmic-dynein
#10
Feng J Gao, Liang Shi, Timothy Hines, Sachin Hebbar, Kristi L Neufeld, Deanna S Smith
Diabetes is linked to an increased risk for colorectal cancer, but the mechanistic underpinnings of this clinically important effect are unclear. Here we describe an interaction between the microtubule motor cytoplasmic dynein, the adenomatous polyposis coli tumor suppressor protein (APC), and glycogen synthase kinase-3β (GSK-3β) that could shed light on this issue. GSK-3β is perhaps best known for glycogen regulation, being inhibited downstream in an insulin-signaling pathway. However, the kinase is also important in many other processes...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057764/regulation-of-yeast-escrt-iii-membrane-scission-activity-by-the-doa4-ubiquitin-hydrolase
#11
Natalie Johnson, Matt West, Greg Odorizzi
ESCRT-III executes membrane scission during the budding of intralumenal vesicles (ILVs) at endosomes. The scission mechanism is unknown but appears to be linked to the cycle of assembly and disassembly of ESCRT-III complexes at membranes. Regulating this cycle is, therefore, expected to be important for determining the timing of ESCRT-III-mediated membrane scission. We show in Saccharomyces cerevisiae that ESCRT-III complexes are stabilized and ILV membrane scission is delayed by Doa4, which is the ubiquitin hydrolase that deubiquitinates transmembrane proteins sorted as cargoes into ILVs...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057763/the-ultrastructural-organization-of-actin-and-myosin-ii-filaments-in-the-contractile-ring-new-support-for-an-old-model-of-cytokinesis
#12
John H Henson, Casey E Ditzler, Aphnie Germain, Patrick M Irwin, Eric T Vogt, Shucheng Yang, Xufeng Wu, Charles B Shuster
Despite recent advances in our understanding of the components and spatial regulation of the contractile ring (CR), the precise ultrastructure of actin and myosin II within the animal cell CR remains an unanswered question. We employed super-resolution light microscopy and platinum replica transmission electron microscopy (TEM) to determine the structural organization of actin and myosin II in isolated cortical cytoskeletons prepared from dividing sea urchin embryos. 3D structured illumination microscopy (SIM) indicated that within the CR, actin and myosin II filaments were organized into tightly packed linear arrays oriented along the axis of constriction, and restricted to a narrow zone within the furrow...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057762/selection-and-stabilization-of-endocytic-sites-by-ede1-a-yeast-functional-homolog-of-the-human-eps15
#13
Rebecca Lu, David G Drubin
During clathrin-mediated endocytosis (CME), endocytic site maturation can be divided into two stages corresponding to the arrival of the early and the late proteins at the plasma membrane. The early proteins are required to capture cargo and to position the late machinery, which includes proteins involved in actin assembly and membrane scission. However, the mechanism by which early arriving proteins select and stabilize endocytic sites is not known. Ede1, one of the earliest proteins recruited to endocytic sites, facilitates site initiation and stabilization...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057761/microtubules-are-reversibly-depolymerized-in-response-to-changing-gaseous-microenvironments-within-aspergillus-nidulans-biofilms
#14
Nandini Shukla, Aysha H Osmani, Stephen A Osmani
How microtubules (MTs) are regulated during fungal biofilm formation is unknown. By tracking MT +end binding proteins (+TIPS) in Aspergillus nidulans we report MTs are regulated to depolymerize within forming fungal biofilms. During this process EB1, dynein and ClipA form transient fibrous then bar-like structures, novel configurations for +TIPS. Cells also respond in an automatous manner with cells separated by a septum able to maintain different MT dynamics. Surprisingly, all cells with depolymerized MTs rapidly repolymerize their MTs after air exchange above the static culture media of biofilms...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28057760/chromatin-and-lamin-a-determine-two-different-mechanical-response-regimes-of-the-cell-nucleus
#15
Andrew D Stephens, Edward J Banigan, Stephen A Adam, Robert D Goldman, John F Marko
The cell nucleus must continually resist and respond to inter- and intracellular mechanical forces to transduce mechanical signals and maintain proper genome organization and expression. Altered nuclear mechanics are associated with many human diseases, including heart disease, progeria, and cancer. Chromatin and nuclear envelope A-type lamin proteins are known to be key nuclear mechanical components perturbed in these diseases, but their distinct mechanical contributions are not known. Here, we directly establish the separate roles of chromatin and lamin A/C and show that they determine two distinct mechanical regimes via micromanipulation of single isolated nuclei...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28031257/sexual-pheromone-modulates-the-frequency-of-cytosolic-ca2-bursts-in-saccharomyces-cerevisiae
#16
Natalia Carbó, Nahuel Tarkowski, Emiliano Perez Ipiña, Silvina Ponce Dawson, Pablo S Aguilar
Transient and highly regulated elevations of cytosolic Ca(2+) control a variety of cellular processes. Bulk measurements using radioactive Ca(2+) and the luminescent sensor aequorin have shown that in response to pheromone, budding yeast cells experience a rise of cytosolic Ca(2+) that is mediated by two import systems composed by the Mid1-Cch1-Ecm7 protein complex, and the Fig 1 protein. Although this response has been largely studied, there is no report on Ca(2+) dynamics at the single cell level. Here, using protein calcium indicators we show that both vegetative and pheromone-treated yeast cells exhibit discrete and asynchronous Ca(2+) bursts...
December 28, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27974642/the-nanoscale-spatial-organization-of-b-cell-receptors-on-igm-and-igg-expressing-human-b-cells
#17
Jinmin Lee, Prabuddha Sengupta, Joseph Brzostowski, Jennifer Lippincott-Schwartz, Susan K Pierce
B cell activation is initiated by the binding of antigen to the B cell receptor (BCR). Here we used dSTORM super resolution imaging to characterize the nanoscale spatial organization of IgM and IgG BCRs on the surfaces of resting and antigen-activated human peripheral blood B cells. We provide insights into both the fundamental process of antigen-driven BCR clustering as well as differences in the spatial organization of IgM and IgG BCRs that may contribute to the characteristic differences in the responses of naïve and memory B cells to antigen...
December 14, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27974641/proper-cytoskeletal-architecture-beneath-the-plasma-membrane-of-red-blood-cells-requires-ttll4
#18
Faryal Ijaz, Yasue Hatanaka, Takahiro Hatanaka, Koji Tsutsumi, Takayuki Iwaki, Kazuo Umemura, Koji Ikegami, Mitsutoshi Setou
Mammalian red blood cells (RBCs) circulate through blood vessels, including capillaries, for tens of days under high mechanical stress. RBCs tolerate this mechanical stress while maintaining their shape because of their elastic membrane skeleton. This membrane skeleton consists of spectrin-actin lattices arranged as quasi-hexagonal units beneath the plasma membrane. In this study, we found that the organization of the RBC cytoskeleton requires tubulin tyrosine ligase-like 4 (Ttll4). RBCs from Ttll4-knockout mice showed larger average diameters in smear tests...
December 14, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27974640/fidgetin-regulates-cultured-astrocyte-migration-by-severing-tyrosinated-microtubules-at-the-leading-edge
#19
Zunlu Hu, Jie Feng, Weijuan Bo, Ronghua Wu, Zhangji Dong, Yan Liu, Liang Qiang, Mei Liu
Microtubule (MT) organization is essential for many cellular events including mitosis, migration and cell polarity. Fidgetin (Fign), an ATP-dependent MT-severing protein, contributes to the regulation of MT configuration by cutting and trimming MT polymers. Functions of Fign have been indicated in neurite outgrowth, mitosis, meiosis, and cellular migration. Here we focused on migration of astrocytes. We report that Fign plays an essential role in cultured astrocyte migration by preferentially targeting MTs (or regions of MTs) that are rich in tyrosinated tubulin, a marker for especially dynamic MTs or especially dynamic regions of MTs...
December 14, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27974639/visualizing-the-dynamic-coupling-of-claudin-strands-to-the-actin-cytoskeleton-through-zo-1
#20
Christina M Van Itallie, Amber Jean Tietgens, James M Anderson
The organization and integrity of epithelial tight junctions depend on interactions between claudins, ZO scaffolding proteins and the cytoskeleton. However, although binding between claudins and ZO-1/2/3 and between ZO-1/2/3 and numerous cytoskeletal proteins has been demonstrated in vitro, FRAP (Fluorescence Recovery After Photobleaching) analysis suggests interactions in vivo are likely highly dynamic. Here we use super resolution live cell imaging in a model fibroblast system to examine relationships between claudins, ZO-1, occludin and actin...
December 14, 2016: Molecular Biology of the Cell
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