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https://www.readbyqxmd.com/read/28194011/self-organizing-actin-patterns-shape-membrane-architecture-but-not-cell-mechanics
#1
M Fritzsche, D Li, H Colin-York, V T Chang, E Moeendarbary, J H Felce, E Sezgin, G Charras, E Betzig, C Eggeling
Cell-free studies have demonstrated how collective action of actin-associated proteins can organize actin filaments into dynamic patterns, such as vortices, asters and stars. Using complementary microscopic techniques, we here show evidence of such self-organization of the actin cortex in living HeLa cells. During cell adhesion, an active multistage process naturally leads to pattern transitions from actin vortices over stars into asters. This process is primarily driven by Arp2/3 complex nucleation, but not by myosin motors, which is in contrast to what has been theoretically predicted and observed in vitro...
February 13, 2017: Nature Communications
https://www.readbyqxmd.com/read/28186323/the-arp2-3-inhibitory-protein-arpin-is-dispensable-for-chemotaxis
#2
Irène Dang, Joern Linkner, Jun Yan, Daniel Irimia, Jan Faix, Alexis Gautreau
Arpin is an Arp2/3 inhibitory protein, which decreases the protrusion lifetime and hence directional persistence in the migration of diverse cells. Arpin is activated by the small GTPase Rac, which controls cell protrusion, thus closing a negative feedback loop that renders the protrusion intrinsically unstable. Because of these properties, it was proposed that Arpin might play a role in directed migration, where directional persistence has to be fine-tuned. We report here, however, that Arpin depleted tumor cells and Arpin knock-out Dictyostelium amoeba display no obvious defect in chemotaxis...
February 10, 2017: Biology of the Cell
https://www.readbyqxmd.com/read/28161632/mammalian-verprolin-cr16-acts-as-a-modulator-of-itsn-scaffold-proteins-association-with-actin
#3
Sergii Kropyvko, Tetyana Gryaznova, Dmytro Morderer, Alla Rynditch
Actin cytoskeleton rearrangements are required for normal cell functioning, and their deregulation leads to various pathologies. Members of two mammalian protein families - ITSNs (ITSN1 and ITSN2) and verprolins (WIP, CR16 and WIRE) are involved in Cdc42/N-WASP/Arp2/3 signaling pathway-mediated remodeling of the actin cytoskeleton. Recently we demonstrated that ITSNs interact with the actin-regulating protein WIP. Here, we show that other member of verprolin family, CR16, also forms complexes with ITSN1 and ITSN2 in human cell lines...
February 1, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28154159/the-t-cell-ift20-interactome-reveals-new-players-in-immune-synapse-assembly
#4
Donatella Galgano, Anna Onnis, Elisa Pappalardo, Federico Galvagni, Oreste Acuto, Cosima T Baldari
Sustained signalling at the immune synapse (IS) requires the synaptic delivery of recycling endosome-associated TCRs. IFT20, a component of the intraflagellar transport system, controls TCR recycling to the IS as a complex with IFT57 and IFT88. Here, we used quantitative mass spectrometry to identify additional interaction partners of IFT20 in Jurkat T cells. In addition to IFT57 and IFT88, the analysis revealed new binding partners, including IFT54, GMAP-210, Arp2/3 complex subunit-3 (ARPC3), COP9 signalosome subunit-1 (CSN1), and ERGIC-53...
February 2, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28128308/%C3%AE-v-class-integrins-exert-dual-roles-on-%C3%AE-5%C3%AE-1-integrins-to-strengthen-adhesion-to-fibronectin
#5
Mitasha Bharadwaj, Nico Strohmeyer, Georgina P Colo, Jonne Helenius, Niko Beerenwinkel, Herbert B Schiller, Reinhard Fässler, Daniel J Müller
Upon binding to the extracellular matrix protein, fibronectin, αV-class and α5β1 integrins trigger the recruitment of large protein assemblies and strengthen cell adhesion. Both integrin classes have been functionally specified, however their specific roles in immediate phases of cell attachment remain uncharacterized. Here, we quantify the adhesion of αV-class and/or α5β1 integrins expressing fibroblasts initiating attachment to fibronectin (≤120 s) by single-cell force spectroscopy. Our data reveals that αV-class integrins outcompete α5β1 integrins...
January 27, 2017: Nature Communications
https://www.readbyqxmd.com/read/28115704/wave1-in-neurons-expressing-the-d1-dopamine-receptor-regulates-cellular-and-behavioral-actions-of-cocaine
#6
Ilaria Ceglia, Ko-Woon Lee, Michael E Cahill, Steven M Graves, David Dietz, Dalton J Surmeier, Eric J Nestler, Angus C Nairn, Paul Greengard, Yong Kim
Wiskott-Aldrich syndrome protein (WASP) family verprolin homologous protein 1 (WAVE1) regulates actin-related protein 2/3 (Arp2/3) complex-mediated actin polymerization. Our previous studies have found WAVE1 to be inhibited by Cdk5-mediated phosphorylation in brain and to play a role in the regulation of dendritic spine morphology. Here we report that mice in which WAVE1 was knocked out (KO) in neurons expressing the D1 dopamine receptor (D1-KO), but not mice where WAVE1 was knocked out in neurons expressing the D2 dopamine receptor (D2-KO), exhibited a significant decrease in place preference associated with cocaine...
February 7, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28106075/phosphatidylinositol-3-4-bisphosphate-regulates-neurite-initiation-and-dendrite-morphogenesis-via-actin-aggregation
#7
Shu-Xin Zhang, Li-Hui Duan, Shun-Ji He, Gui-Feng Zhuang, Xiang Yu
Neurite initiation is critical for neuronal morphogenesis and early neural circuit development. Recent studies showed that local actin aggregation underneath the cell membrane determined the site of neurite initiation. An immediately arising question is what signaling mechanism initiated actin aggregation. Here we demonstrate that local clustering of phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2), a phospholipid with relatively few known signaling functions, is necessary and sufficient for aggregating actin and promoting neuritogenesis...
January 20, 2017: Cell Research
https://www.readbyqxmd.com/read/28059163/phosphoproteomic-comparison-of-pik3ca-and-pten-signalling-identifies-the-nucleotidase-nt5c-as-a-novel-akt-substrate
#8
Larissa S Moniz, Silvia Surinova, Essam Ghazaly, Lorena Gonzalez Velasco, Syed Haider, Juan Carlos Rodríguez-Prados, Inma M Berenjeno, Claude Chelala, Bart Vanhaesebroeck
To identify novel effectors and processes regulated by PI3K pathway activation, we performed an unbiased phosphoproteomic screen comparing two common events of PI3K deregulation in cancer: oncogenic Pik3ca mutation (Pik3ca(H1047R)) and deletion of Pten. Using mouse embryonic fibroblast (MEF) models that generate inducible, low-level pathway activation as observed in cancer, we quantified 7566 unique phosphopeptides from 3279 proteins. A number of proteins were found to be differentially-regulated by Pik3ca(H1047R) and Pten loss, suggesting unique roles for these two events in processes such as vesicular trafficking, DNA damage repair and RNA splicing...
January 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28013022/nano-scale-actin-network-characterization-of-fibroblast-cells-lacking-functional-arp2-3-complex
#9
Karen L Anderson, Christopher Page, Mark F Swift, Praveen Suraneni, Mandy E W Janssen, Thomas D Pollard, Rong Li, Niels Volkmann, Dorit Hanein
Arp2/3 complex is thought to be the primary protrusive force generator in cell migration by controlling the assembly and turnover of the branched filament network that pushes the leading edge of moving cells forward. However, mouse fibroblasts without functional Arp2/3 complex migrate at rates similar to wild-type cells, contradicting this paradigm. We show by correlative fluorescence and large-scale cryo-tomography studies combined with automated actin-network analysis that the absence of functional Arp2/3 complex has profound effects on the nano-scale architecture of actin networks...
December 21, 2016: Journal of Structural Biology
https://www.readbyqxmd.com/read/27998771/deficiency-in-ubiquitin-like-protein-ubl4a-impairs-migration-of-fibroblasts-and-macrophages
#10
Yu Zhao, Huaiyuan Zhang, Carlos Affonso, Raiza Bonomo, Adriana Mañas, Jialing Xiang
Ubiquitin-like protein Ubl4A is a small, multi-functional protein with no ubiquitination activity. We have previously demonstrated that Ubl4A directly interacts with actin-related protein 2/3 complex (Arp2/3) and promotes Arp2/3-dependent actin branching, thereby accelerating plasma membrane translocation of protein kinase Akt upon insulin stimulation. Here, we show that Ubl4A is critical for plasma membrane protrusion and cell migration. Ubl4A, F-actin and Arp2/3 are co-localized at the cell leading edges during wound closure...
January 29, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27998058/label-free-neuroproteomics-of-the-hippocampal-accumbal-circuit-reveals-deficits-in-neurotransmitter-and-neuropeptide-signaling-in-mice-lacking-ethanol-sensitive-adenosine-transporter
#11
Alfredo Oliveros, Phillip Starski, Daniel Lindberg, Sun Choi, Carrie J Heppelmann, Surendra Dasari, Doo-Sup Choi
The neural circuit of the dorsal hippocampus (dHip) and nucleus accumbens (NAc) contributes to cue-induced learning and addictive behaviors, as demonstrated by the escalation of ethanol-seeking behaviors observed following deletion of the adenosine equilibrative nucleoside transporter 1 (ENT1-/-) in mice. Here, we perform a quantitative LC-MS/MS neuroproteomics in the dHip and NAc of ENT1-/- mice. Using Ingenuity Pathway Analysis, we identified proteins associated with increased long-term potentiation, ARP2/3-mediated actin cytoskeleton signaling...
December 20, 2016: Journal of Proteome Research
https://www.readbyqxmd.com/read/27997888/rasd1-knockdown-results-in-failure-of-oocyte-maturation
#12
Youngeun Lee, Kyeoung-Hwa Kim, Hyemin Yoon, Ok-Hee Lee, Eunyoung Kim, Miseon Park, Hoon Jang, Kwonho Hong, Hyuk Song, Jung Jae Ko, Woo Sik Lee, Kyung-Ah Lee, Eun Mi Chang, Youngsok Choi
BACKGROUND: Ras dexamethasone-induced protein (RASD1) is a member of Ras superfamily of small GTPases. RASD1 regulates various signaling pathways involved in iron homeostasis, growth hormone secretion, and circadian rhythm. However, RASD1 function in oocyte remains unknown. METHODS: Using immunohistochemistry, immunofluorescence, and quantitative real-time RT-PCR, RASD1 expression in mouse ovary and RASD1 role in oocyte maturation-related gene expression, spindle formation, and chromosome alignment were analyzed...
2016: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/27957528/arp2-3-and-vasp-are-essential-for-fear-memory-formation-in-lateral-amygdala
#13
Sreetama Basu, Irina Kustanovich, Raphael Lamprecht
The actin cytoskeleton is involved in key neuronal functions such as synaptic transmission and morphogenesis. However, the roles and regulation of actin cytoskeleton in memory formation remain to be clarified. In this study, we unveil the mechanism whereby actin cytoskeleton is regulated to form memory by exploring the roles of the major actin-regulatory proteins Arp2/3, VASP, and formins in long-term memory formation. Inhibition of Arp2/3, involved in actin filament branching and neuronal morphogenesis, in lateral amygdala (LA) with the specific inhibitor CK-666 during fear conditioning impaired long-term, but not short-term, fear memory...
November 2016: ENeuro
https://www.readbyqxmd.com/read/27939292/structural-basis-of-arp2-3-complex-inhibition-by-gmf-coronin-and-arpin
#14
Olga S Sokolova, Angelina Chemeris, Siyang Guo, Salvatore L Alioto, Meghal Gandhi, Shae Padrick, Evgeniya Pechnikova, Violaine David, Alexis Gautreau, Bruce L Goode
The evolutionarily conserved Arp2/3 complex plays a central role in nucleating the branched actin filament arrays that drive cell migration, endocytosis, and other processes. To better understand Arp2/3 complex regulation, we used single-particle electron microscopy to compare the structures of Arp2/3 complex bound to three different inhibitory ligands: glia maturation factor (GMF), Coronin, and Arpin. Although the three inhibitors have distinct binding sites on Arp2/3 complex, they each induced an "open" nucleation-inactive conformation...
January 20, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/27927957/the-proteome-of-bloc-1-genetic-defects-identifies-the-arp2-3-actin-polymerization-complex-to-function-downstream-of-the-schizophrenia-susceptibility-factor-dysbindin-at-the-synapse
#15
Avanti Gokhale, Cortnie Hartwig, Amanda H Freeman, Ravi Das, Stephanie A Zlatic, Rachel Vistein, Amelia Burch, Guillemette Carrot, Arielle F Lewis, Sheldon Nelms, Dion K Dickman, Manojkumar A Puthenveedu, Daniel N Cox, Victor Faundez
: Proteome modifications downstream of monogenic or polygenic disorders have the potential to uncover novel molecular mechanisms participating in pathogenesis and/or extragenic modification of phenotypic expression. We tested this idea by determining the proteome sensitive to genetic defects in a locus encoding dysbindin, a protein required for synapse biology and implicated in schizophrenia risk. We applied quantitative mass spectrometry to identify proteins expressed in neuronal cells the abundance of which was altered after downregulation of the schizophrenia susceptibility factor dysbindin (Bloc1s8) or two other dysbindin-interacting polypeptides, which assemble into the octameric biogenesis of lysosome-related organelles complex 1 (BLOC-1)...
December 7, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/27924678/a-novel-role-for-14-kda-phosphohistidine-phosphatase-in-lamellipodia-formation
#16
Anjian Xu, Xiaojin Li, Siwen Li, Lan Sun, Shanna Wu, Bei Zhang, Jian Huang
Cell migration involves dynamic regulation of the actin cytoskeleton, which exhibits rapid actin polymerization at the leading edge of migrating cells. This process relies on regulated recruitment of actin nucleators and actin-binding proteins to the leading edge to polymerize new actin filaments. Many of these proteins have been identified, including the actin-related protein (Arp) 2/3 complex, which has emerged as the core player in the initiation of actin polymerization. However, the functional coordination of these proteins is unclear...
December 7, 2016: Cell Adhesion & Migration
https://www.readbyqxmd.com/read/27922825/model-of-turnover-kinetics-in-the-lamellipodium-implications-of-slow-and-fast-diffusing-capping-protein-and-arp2-3-complex
#17
Laura M McMillen, Dimitrios Vavylonis
Cell protrusion through polymerization of actin filaments at the leading edge of motile cells may be influenced by spatial gradients of diffuse actin and regulators. Here we study the distribution of two of the most important regulators, capping protein and Arp2/3 complex, which regulate actin polymerization in the lamellipodium through capping and nucleation of free barbed ends. We modeled their kinetics using data from prior single molecule microscopy experiments on XTC cells. These experiments have provided evidence for a broad distribution of diffusion coefficients of both capping protein and Arp2/3 complex...
December 6, 2016: Physical Biology
https://www.readbyqxmd.com/read/27911716/wasp-family-proteins-more-than-arp2-3-activators
#18
REVIEW
Joe J Tyler, Ellen G Allwood, Kathryn R Ayscough
Wiskott-Aldrich syndrome protein (WASP) family proteins have been extensively characterized as factors that promote the nucleation of actin through the activation of the protein complex Arp2/3. While yeast mostly have a single member of the family, mammalian cells have at least six different members, often with multiple isoforms. Members of the family are characterized by a common structure. Their N-termini are varied and are considered to confer spatial and temporal regulation of Arp2/3-activating activity, whereas their C-terminal half contains a polyproline-rich region, one or more WASP homology-2 (WH2) actin-binding domains and motifs that bind directly to Arp2/3...
October 15, 2016: Biochemical Society Transactions
https://www.readbyqxmd.com/read/27904039/role-of-rac1-wave2-signaling-in-mediating-the-inhibitory-effects-of-%C3%AE-tocotrienol-on-mammary-cancer-cell-migration-and-invasion
#19
Ibrahim Gayadh Algayadh, Venkateshwararao Dronamraju, Paul William Sylvester
The majority of breast cancer deaths result from the progression of this disease to a metastatic phenotype. Rac1 and Cdc42 are Rho family members that together with their downstream effectors, Wiskott-Aldrich Syndrome protein-family verprolin-homologous protein 2 (WAVE2) and Arp2/3, play an important role in cytoskeletal reorganization and the formation of membrane protrusions that promote cancer cell migration and invasion. γ-Tocotrienol, is a natural isoform within the vitamin E family of compounds that inhibits breast cancer cell growth and progression by suppressing various signaling pathways involved in mitogenic signaling and metastatic progression...
2016: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/27900558/the-role-of-viral-protein-ac34-in-nuclear-relocation-of-subunits-of-the-actin-related-protein-2-3-complex
#20
Jingfang Mu, Yongli Zhang, Yangyang Hu, Xue Hu, Yuan Zhou, Xinwen Chen, Yun Wang
The actin nucleator actin-related protein complex (Arp2/3) is composed of seven subunits: Arp2, Arp3, p40/ARPC1 (P40), p34/ARPC2 (P34), p21/ARPC3 (P21), p20/ARPC4 (P20), and p16/ARPC5 (P16). Arp2/3 plays crucial roles in a variety of cellular activities through regulation of actin polymerization. Autographa californica multiple nucleopolyhedrovirus (AcMNPV), one of the beststudied alphabaculoviruses, induces Arp2/3 nuclear relocation and mediates nuclear actin polymerization to assist in virus replication. We have demonstrated that Ac34, a viral late-gene product, induces translocation of the P40 subunit of Arp2/3 to the nucleus during AcMNPV infection...
December 2016: Virologica Sinica
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