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https://www.readbyqxmd.com/read/27440924/formin-mediated-actin-polymerization-at-cell-cell-junctions-stabilizes-e-cadherin-and-maintains-monolayer-integrity-during-wound-repair
#1
Megha Vaman Rao, Ronen Zaidel-Bar
Cadherin-mediated cell-cell adhesion is required for epithelial tissue integrity in homeostasis, during development, and in tissue repair. E-cadherin stability depends on F-actin, but the mechanisms regulating actin polymerization at cell-cell junctions remain poorly understood. Here we investigated a role for formin-mediated actin polymerization at cell-cell junctions. We identify mDia1 and Fmnl3 as major factors enhancing actin polymerization and stabilizing E-cadherin at epithelial junctions. Fmnl3 localizes to adherens junctions downstream of Src and Cdc42 and its depletion leads to a reduction in F-actin and E-cadherin at junctions and a weakening of cell-cell adhesion...
September 15, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27378434/pfa-fixation-enables-artifact-free-super-resolution-imaging-of-the-actin-cytoskeleton-and-associated-proteins
#2
Daniela Leyton-Puig, Katarzyna M Kedziora, Tadamoto Isogai, Bram van den Broek, Kees Jalink, Metello Innocenti
Super-resolution microscopy (SRM) allows precise localization of proteins in cellular organelles and structures, including the actin cytoskeleton. Yet sample preparation protocols for SRM are rather anecdotal and still being optimized. Thus, SRM-based imaging of the actin cytoskeleton and associated proteins often remains challenging and poorly reproducible. Here, we show that proper paraformaldehyde (PFA)-based sample preparation preserves the architecture of the actin cytoskeleton almost as faithfully as gold-standard glutaraldehyde fixation...
2016: Biology Open
https://www.readbyqxmd.com/read/27199431/accelerated-actin-filament-polymerization-from-microtubule-plus-ends
#3
Jessica L Henty-Ridilla, Aneliya Rankova, Julian A Eskin, Katelyn Kenny, Bruce L Goode
Microtubules (MTs) govern actin network remodeling in a wide range of biological processes, yet the mechanisms underlying this cytoskeletal cross-talk have remained obscure. We used single-molecule fluorescence microscopy to show that the MT plus-end-associated protein CLIP-170 binds tightly to formins to accelerate actin filament elongation. Furthermore, we observed mDia1 dimers and CLIP-170 dimers cotracking growing filament ends for several minutes. CLIP-170-mDia1 complexes promoted actin polymerization ~18 times faster than free-barbed-end growth while simultaneously enhancing protection from capping proteins...
May 20, 2016: Science
https://www.readbyqxmd.com/read/27177153/matrix-metalloproteinase%C3%A2-2-regulates-mda%C3%A2-mb%C3%A2-231-breast-cancer-cell-invasion-induced-by-active-mammalian-diaphanous-related-formin%C3%A2-1
#4
Daehwan Kim, Sangmyung Rhee
Mammalian diaphanous‑related formin 1 (mDia1) was initially identified as a Rho GTPase effector involved in the progression of various diseases, including types of cancer. However, the precise underlying molecular mechanism of mDia1‑mediated cancer cell invasion remains to be elucidated. In the present study, mDia1 expression was demonstrated to be upregulated in tissues from a number of cancer types, including kidney, prostate, and breast cancer using immunohistochemical analysis. Forced expression of a constitutively active (CA) form of mDia1 induces invasion, as measured by Transwell invasion assay, of MDA‑MB‑231 cells, which is a highly invasive breast cancer cell line, and this effect was markedly impaired by matrix metalloproteinase (MMP)‑2 silencing...
July 2016: Molecular Medicine Reports
https://www.readbyqxmd.com/read/27030671/an-mdia1-inf2-formin-activation-cascade-facilitated-by-iqgap1-regulates-stable-microtubules-in-migrating-cells
#5
Francesca Bartolini, Laura Andres-Delgado, Xiaoyi Qu, Sara Nik, Nagendran Ramalingam, Lenor Kremer, Miguel A Alonso, Gregg G Gundersen
Multiple formins regulate microtubule (MT) arrays, but whether they function individually or in a common pathway is unknown. Lysophosphatidic acid (LPA) stimulates the formation of stabilized detyrosinated MTs (Glu MTs) in NIH3T3 fibroblasts through RhoA and the formin mDia1. Here we show that another formin, INF2, is necessary for mDia1-mediated induction of Glu MTs and regulation of MT dynamics and that mDia1 can be bypassed by activating INF2. INF2 localized to MTs after LPA treatment in an mDia1-dependent manner, suggesting that mDia1 regulates INF2...
June 1, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/26912794/mechanical-confinement-triggers-glioma-linear-migration-dependent-on-formin-fhod3
#6
Pascale Monzo, Yuk Kien Chong, Charlotte Guetta-Terrier, Anitha Krishnasamy, Sharvari R Sathe, Evelyn K F Yim, Wai Hoe Ng, Beng Ti Ang, Carol Tang, Benoit Ladoux, Nils C Gauthier, Michael P Sheetz
Glioblastomas are extremely aggressive brain tumors with highly invasive properties. Brain linear tracks such as blood vessel walls constitute their main invasive routes. Here we analyze rat C6 and patient-derived glioma cell motility in vitro using micropatterned linear tracks to mimic blood vessels. On laminin-coated tracks (3-10 μm), these cells used an efficient saltatory mode of migration similar to their in vivo migration. This saltatory migration was also observed on larger tracks (50-400 μm in width) at high cell densities...
April 15, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/26898799/smifh2-mediated-mdia-formin-functional-inhibition-potentiates-chemotherapeutic-targeting-of-human-ovarian-cancer-spheroids
#7
Megan A Ziske, Krista M Pettee, MaNada Khaing, Kaitlin Rubinic, Kathryn M Eisenmann
Due to a lack of effective screening or prevention protocol for epithelial ovarian cancer (EOC), there is a critical unmet need to develop therapeutic interventions for EOC treatment. EOC metastasis is unique. Initial dissemination is not primarily hematogenous, yet is facilitated through shedding of primary tumor cells into the peritoneal fluid and accumulating ascites. Increasingly, isolated patient spheroids point to a clinical role for spheroids in EOC metastasis. EOC spheroids are highly invasive structures that disseminate upon peritoneal mesothelium, and visceral tissues including liver and omentum...
March 25, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/26893363/mdia1-regulates-breast-cancer-invasion-by-controlling-membrane-type-1-matrix-metalloproteinase-localization
#8
Daehwan Kim, Jangho Jung, Eunae You, Panseon Ko, Somi Oh, Sangmyung Rhee
Mammalian diaphanous-related formin 1 (mDia1) expression has been linked with progression of malignant cancers in various tissues. However, the precise molecular mechanism underlying mDia1-mediated invasion in cancer cells has not been fully elucidated. In this study, we found that mDia1 is upregulated in invasive breast cancer cells. Knockdown of mDia1 in invasive breast cancer profoundly reduced invasive activity by controlling cellular localization of membrane type 1-matrix metalloproteinase (MT1-MMP) through interaction with microtubule tracks...
April 5, 2016: Oncotarget
https://www.readbyqxmd.com/read/26674774/coxiella-burnetii-phagocytosis-is-regulated-by-gtpases-of-the-rho-family-and-the-rhoa-effectors-mdia1-and-rock
#9
Romina P Salinas, Rodolfo M Ortiz Flores, Jesús S Distel, Milton O Aguilera, María I Colombo, Walter Berón
The GTPases belonging to the Rho family control the actin cytoskeleton rearrangements needed for particle internalization during phagocytosis. ROCK and mDia1 are downstream effectors of RhoA, a GTPase involved in that process. Coxiella burnetii, the etiologic agent of Q fever, is internalized by the host´s cells in an actin-dependent manner. Nevertheless, the molecular mechanism involved in this process has been poorly characterized. This work analyzes the role of different GTPases of the Rho family and some downstream effectors in the internalization of C...
2015: PloS One
https://www.readbyqxmd.com/read/26641718/innate-control-of-actin-nucleation-determines-two-distinct-migration-behaviours-in-dendritic-cells
#10
Pablo Vargas, Paolo Maiuri, Marine Bretou, Pablo J Sáez, Paolo Pierobon, Mathieu Maurin, Mélanie Chabaud, Danielle Lankar, Dorian Obino, Emmanuel Terriac, Matthew Raab, Hawa-Racine Thiam, Thomas Brocker, Susan M Kitchen-Goosen, Arthur S Alberts, Praveen Sunareni, Sheng Xia, Rong Li, Raphael Voituriez, Matthieu Piel, Ana-Maria Lennon-Duménil
Dendritic cell (DC) migration in peripheral tissues serves two main functions: antigen sampling by immature DCs, and chemokine-guided migration towards lymphatic vessels (LVs) on maturation. These migratory events determine the efficiency of the adaptive immune response. Their regulation by the core cell locomotion machinery has not been determined. Here, we show that the migration of immature DCs depends on two main actin pools: a RhoA-mDia1-dependent actin pool located at their rear, which facilitates forward locomotion; and a Cdc42-Arp2/3-dependent actin pool present at their front, which limits migration but promotes antigen capture...
January 2016: Nature Cell Biology
https://www.readbyqxmd.com/read/26609804/interaction-between-mdia1-and-rock-in-rho-induced-migration-and-adhesion-of-human-dental-pulp-cells
#11
L Cheng, J Xu, Y Y Qian, H Y Pan, H Yang, M Y Shao, R Cheng, T Hu
AIM: To investigate the effects of mammalian homolog of Drosophila diaphanous-1(mDia1) and Rho-associated coiled-coil-containing protein kinase (ROCK) on the migration and adhesion of dental pulp cells (DPCs). METHODOLOGY: Lysophosphatidic acid (LPA) was used to activate Rho signaling. mDia1 and ROCK were inhibited by short interfering RNA and the specific inhibitor, Y-27632, respectively. The migration of DPCs was assessed using the transwell migration assay and scratch test...
November 26, 2015: International Endodontic Journal
https://www.readbyqxmd.com/read/26566078/single-molecule-visualization-of-a-formin-capping-protein-decision-complex-at-the-actin-filament-barbed-end
#12
Jeffrey P Bombardier, Julian A Eskin, Richa Jaiswal, Ivan R Corrêa, Ming-Qun Xu, Bruce L Goode, Jeff Gelles
Precise control of actin filament length is essential to many cellular processes. Formins processively elongate filaments, whereas capping protein (CP) binds to barbed ends and arrests polymerization. While genetic and biochemical evidence has indicated that these two proteins function antagonistically, the mechanism underlying the antagonism has remained unresolved. Here we use multi-wavelength single-molecule fluorescence microscopy to observe the fully reversible formation of a long-lived 'decision complex' in which a CP dimer and a dimer of the formin mDia1 simultaneously bind the barbed end...
November 13, 2015: Nature Communications
https://www.readbyqxmd.com/read/26564775/formin-and-capping-protein-together-embrace-the-actin-filament-in-a-m%C3%A3-nage-%C3%A3-trois
#13
Shashank Shekhar, Mikael Kerleau, Sonja Kühn, Julien Pernier, Guillaume Romet-Lemonne, Antoine Jégou, Marie-France Carlier
Proteins targeting actin filament barbed ends play a pivotal role in motile processes. While formins enhance filament assembly, capping protein (CP) blocks polymerization. On their own, they both bind barbed ends with high affinity and very slow dissociation. Their barbed-end binding is thought to be mutually exclusive. CP has recently been shown to be present in filopodia and controls their morphology and dynamics. Here we explore how CP and formins may functionally coregulate filament barbed-end assembly...
2015: Nature Communications
https://www.readbyqxmd.com/read/26529665/hydrogen-rich-medium-ameliorates-lipopolysaccharide-induced-barrier-dysfunction-via-rhoa-mdia1-signaling-in-caco-2-cells
#14
Tao Yang, Lu Wang, Ruiqiang Sun, Hongguang Chen, Hongtao Zhang, Yang Yu, Yanyan Wang, Guolin Wang, Yonghao Yu, Keliang Xie
Gastrointestinal barrier dysfunction is associated with the severity and prognosis of sepsis. Hydrogen gas (H2) can ameliorate multiple organ damage in septic animals. Ras homolog gene family member A (RhoA) and mammalian diaphanous-related formin 1 (mDia1) are important to regulate tight junction (TJ) and adherens junction (AJ), both of which determine the integrity of the intestinal barrier. This study was aimed to investigate whether H2 could modulate lipopolysaccharide (LPS)-stimulated dysfunction of the intestinal barrier and whether RhoA-mDia1 signaling is involved...
February 2016: Shock
https://www.readbyqxmd.com/read/26446836/cell-type-dependent-mechanisms-for-formin-mediated-assembly-of-filopodia
#15
Lorna E Young, Ernest G Heimsath, Henry N Higgs
Filopodia are finger-like protrusions from the plasma membrane and are of fundamental importance to cellular physiology, but the mechanisms governing their assembly are still in question. One model, called convergent elongation, proposes that filopodia arise from Arp2/3 complex-nucleated dendritic actin networks, with factors such as formins elongating these filaments into filopodia. We test this model using constitutively active constructs of two formins, FMNL3 and mDia2. Surprisingly, filopodial assembly requirements differ between suspension and adherent cells...
December 15, 2015: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/26415699/a-resilient-formin-derived-cortical-actin-meshwork-in-the-rear-drives-actomyosin-based-motility-in-2d-confinement
#16
Nagendran Ramalingam, Christof Franke, Evelin Jaschinski, Moritz Winterhoff, Yao Lu, Stefan Brühmann, Alexander Junemann, Helena Meier, Angelika A Noegel, Igor Weber, Hongxia Zhao, Rudolf Merkel, Michael Schleicher, Jan Faix
Cell migration is driven by the establishment of disparity between the cortical properties of the softer front and the more rigid rear allowing front extension and actomyosin-based rear contraction. However, how the cortical actin meshwork in the rear is generated remains elusive. Here we identify the mDia1-like formin A (ForA) from Dictyostelium discoideum that generates a subset of filaments as the basis of a resilient cortical actin sheath in the rear. Mechanical resistance of this actin compartment is accomplished by actin crosslinkers and IQGAP-related proteins, and is mandatory to withstand the increased contractile forces in response to mechanical stress by impeding unproductive blebbing in the rear, allowing efficient cell migration in two-dimensional-confined environments...
2015: Nature Communications
https://www.readbyqxmd.com/read/26381868/inhibition-of-diaphanous-formin-signaling-in-vivo-impairs-cardiovascular-development-and-alters-smooth-muscle-cell-phenotype
#17
Laura Weise-Cross, Joan M Taylor, Christopher P Mack
OBJECTIVE: We and others have previously shown that RhoA-dependent stimulation of myocardin-related transcription factor nuclear localization promotes smooth muscle cell (SMC) marker gene expression. The goal of this study was to provide direct in vivo evidence that actin polymerization by the diaphanous-related formins contributes to the regulation of SMC differentiation and phenotype. APPROACH AND RESULTS: Conditional Cre-based genetic approaches were used to overexpress a well-characterized dominant-negative variant of mDia1 (DNmDia) in SMC...
November 2015: Arteriosclerosis, Thrombosis, and Vascular Biology
https://www.readbyqxmd.com/read/26349808/initiation-of-lamellipodia-and-ruffles-involves-cooperation-between-mdia1-and-the-arp2-3-complex
#18
Tadamoto Isogai, Rob van der Kammen, Daniela Leyton-Puig, Katarzyna M Kedziora, Kees Jalink, Metello Innocenti
Protrusion of lamellipodia and ruffles requires polymerization of branched actin filaments by the Arp2/3 complex. Although regulation of Arp2/3 complex activity has been extensively investigated, the mechanism of initiation of lamellipodia and ruffles remains poorly understood. Here, we show that mDia1 acts in concert with the Arp2/3 complex to promote initiation of lamellipodia and ruffles. We find that mDia1 is an epidermal growth factor (EGF)-regulated actin nucleator involved in membrane ruffling using a combination of knockdown and rescue experiments...
October 15, 2015: Journal of Cell Science
https://www.readbyqxmd.com/read/26273703/functional-hierarchy-of-redundant-actin-assembly-factors-revealed-by-fine-grained-registration-of-intrinsic-image-fluctuations
#19
Kwonmoo Lee, Hunter L Elliott, Youbean Oak, Chih-Te Zee, Alex Groisman, Jessica D Tytell, Gaudenz Danuser
Highly redundant pathways often contain components whose functions are difficult to decipher from the responses induced by genetic or molecular perturbations. Here, we present a statistical approach that samples and registers events observed in images of intrinsic fluctuations in unperturbed cells to establish the functional hierarchy of events in systems with redundant pathways. We apply this approach to study the recruitment of actin assembly factors involved in the protrusion of the cell membrane. We find that the formin mDia1, along with nascent adhesion components, is recruited to the leading edge of the cell before protrusion onset, initiating linear growth of the lamellipodial network...
July 29, 2015: Cell Systems
https://www.readbyqxmd.com/read/26201082/the-rho-mdia1-signaling-pathway-is-required-for-cyclic-strain-induced-cytoskeletal-rearrangement-of-human-periodontal-ligament-cells
#20
Jiannan Wu, Meng Song, Tengyu Li, Zhengxian Zhu, Jinsong Pan
Tooth movement is the result of periodontal tissue reconstruction. The biomechanical effects produced by orthopedic forces can affect the cytoskeletal rearrangement of human periodontal ligament cells (hPDLCs). However, the mechanisms responsible for the cytoskeletal rearrangement are not completely understood. To analyze the effect, we investigated the role of the Rho-mDia1 signaling pathway in cyclic strain-induced cytoskeletal rearrangement of hPDLCs in detail. We cultured hPDLCs on collagen I-coated six-well Bioflex plates and then exposed them to cyclic strain with physiological loading (10%) at a frequency of 0...
September 10, 2015: Experimental Cell Research
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