Prokaryotic Cytoskeletons: Protein Filaments Organizing Small Cells
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The Cytoskeleton in Cell-Autonomous Immunity: Structural Determinants of Host Defence
Mostowy & Shenoy, Nat Rev Immunol, doi:10.1038/nri3877 The cytoskeleton in cell-autonomous immunity: structural determinants of host defence Serge Mostowy and Avinash R. Shenoy Medical Research Council Centre of Molecular Bacteriology and Infection (CMBI), Imperial College London, Armstrong Road, London SW7 2AZ, UK. e‑mails: [email protected] ; [email protected] doi:10.1038/nri3877 Published online 21 August 2015 Abstract Host cells use antimicrobial proteins, pathogen-restrictive compartmentalization and cell death in their defence against intracellular pathogens. Recent work has revealed that four components of the cytoskeleton — actin, microtubules, intermediate filaments and septins, which are well known for their roles in cell division, shape and movement — have important functions in innate immunity and cellular self-defence. Investigations using cellular and animal models have shown that these cytoskeletal proteins are crucial for sensing bacteria and for mobilizing effector mechanisms to eliminate them. In this Review, we highlight the emerging roles of the cytoskeleton as a structural determinant of cell-autonomous host defence. 1 Mostowy & Shenoy, Nat Rev Immunol, doi:10.1038/nri3877 Cell-autonomous immunity, which is defined as the ability of a host cell to eliminate an invasive infectious agent, is a first line of defence against microbial pathogens 1 . It relies on antimicrobial proteins, specialized degradative compartments and programmed host cell death 1–3 . Cell- autonomous immunity is mediated by tiered innate immune signalling networks that sense microbial pathogens and stimulate downstream pathogen elimination programmes. Recent studies on host– microorganism interactions show that components of the host cell cytoskeleton are integral to the detection of bacterial pathogens as well as to the mobilization of antibacterial responses (FIG. -
How Protein Materials Balance Strength, Robustness and Adaptability
How Protein Materials Balance Strength, Robustness And Adaptability The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Buehler, Markus J., and Yu Ching Yung. “How protein materials balance strength, robustness, and adaptability.” HFSP Journal 4.1 (2010): 26-40. As Published http://dx.doi.org/10.2976/1.3267779 Publisher American Institute of Physics Version Author's final manuscript Citable link http://hdl.handle.net/1721.1/67894 Terms of Use Attribution-Noncommercial-Share Alike 3.0 Unported Detailed Terms http://creativecommons.org/licenses/by-nc-sa/3.0/ How protein materials balance strength, robustness and adaptability Markus J. Buehler1,2,3*, Yu Ching Yung1 1Laboratory for Atomistic and Molecular Mechanics, Department of Civil and Environmental Engineering, 77 Massachusetts Ave. Room 1-235A&B, Cambridge, MA, USA 2Center for Materials Science and Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA, USA 3Center for Computational Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA, USA * E-mail: [email protected], Lab URL: http://web.mit.edu/mbuehler/www/ Abstract: Proteins form the basis of a wide range of biological materials such as hair, skin, bone, spider silk or cells, which play an important role in providing key functions to biological systems. The focus of this article is to discuss how protein materials are capable of balancing multiple, seemingly incompatible properties such as strength, robustness and adaptability. Here we review bottom-up materiomics studies focused on the mechanical behavior of protein materials at multiple scales, from nano to macro. -
Actin Cytoskeleton of Spread Fibroblasts Appears to Assemble at the Cell Edges
J. Cell Sd. 82, 235-248 (1986) 235 Printed in Great Britain © The Company of Biologists Limited 1986 ACTIN CYTOSKELETON OF SPREAD FIBROBLASTS APPEARS TO ASSEMBLE AT THE CELL EDGES TATJANA M. SVITKINA, ALEXANDER A. NEYFAKH, JR Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, Moscow 119899, USSR AND ALEXANDER D. BERSHADSKY All-Union Cancer Research Center, Academy of Medical Sciences, Moscow 115478, USSR SUMMARY The action of metabolic inhibitors on actin cytoskeleton of cultured quail embryo fibroblasts has been studied using electron microscopy of platinum replicas and immunofluorescence microscopy. Sodium azide as well as other inhibitors (oligomycin and dinitrophenol) caused the disassembly of all types of actin structures: actin meshwork at the cell active edges, microfilament sheath underlying the cell surface, and microfilament bundles. Studying the time- and dose-dependence of the destruction process we have found that the active edge meshwork and microfilament sheath are much more labile than microfilament bundles. After the removal of metabolic inhibitors actin cytoskeleton restoration begins at the cell edges. The first sign of this process is the formation of actin meshwork along the whole cell perimeter (l-10min of recovery). Sometimes fragments of this meshwork bend upwards forming ruffles. Later (10-20 min of recovery) the microfilament sheath appears at the cell periphery as a narrow band. The sheath seems to be formed from the edge meshwork, since ruffles in the process of transformation to sheath could be seen. During the following restoration the microfilament sheath gradually expands towards the cell centre. The last step of actin cytoskeleton restoration (60—120 min of recovery) is the formation of bundles. -
HTS-Tubulin Polymerization Assay Biochem Kit™
The Protein Manual Experts Cytoskeleton, Inc. V 8.0 HTS-Tubulin Polymerization Assay Biochem Kit™ (>97% pure tubulin, Porcine Tubulin) Cat. # BK004P Phone: (303) 322.2254 Fax: (303) 322.2257 Customer Service: [email protected] cytoskeleton.com Technical Support: [email protected] cytoskeleton.com Page 2 Manual Contents Section I: Introduction About Tubulin -------------------------------------------------------------------------- 5 About the BK004P polymerization Assay -------------------------------------- 6-7 Comparison of Polymerization Assays ----------------------------------------- 8-9 Section II: Purchaser Notification ------------------------------------------------------------ 10 Section III: Kit Contents ------------------------------------------------------------------------- 11-12 Section V: Reconstitution and Storage of Components ----------------------------- 13 Section IV: Important Technical Notes Notes on Updated version --------------------------------------------------------- 14 Spectrophotometer settings ------------------------------------------------------- 14 Spectrophotometer pathlength---------------------------------------------------- 15 Temperature & time dependence of polymerization ------------------------ 15 Recommended pipetting technique --------------------------------------------- 15-16 Tubulin protein stability ------------------------------------------------------------- 16 Test compound or protein preparation ------------------------------------------ 16-17 Section VI: Assay Protocol -
Essential for Protein Regulation
CYTOSKELETON NEWS NEWS FROM CYTOSKELETON INC. Rac GTP Rho GTPases Control Cell Migration P-Rex1 Related Publications Fli2 Rac Research Tools January GDP 2020 Tiam1 Rac GTP v Rho GTPases Control Cell Migration Meetings Directed cell migration depends upon integrin-containing deposition of the ECM protein fibronectin which supports Cold Spring Harbor focal adhesions connecting the cell’s actin cytoskeleton nascent lamellipodia formation. The fibronectin binds to Conference - Systems with the extracellular matrix (ECM) and transmitting integrin and serves as an ECM substrate for Rac1-dependent 18 Biology: Global Regulation of mechanical force. Focal adhesion formation and subsequent directional migration . N e Gene Expression migration require dynamic re-organization of actin-based w contractile fibers and protrusions at a cell’s trailing and Cell Directionality s March 11-14th leading edges, respectively, in response to extracellular Cold Spring Harbor, NY guidance cues. Migration is essential for healthy cell Rho-family-mediated remodeling of the actin cytoskeleton is Cytoskeleton Supported (and organism) development, growth, maturation, and necessary for the cell’s directional response to extracellular physiological responses to diseases, injuries, and/or immune guidance cues (e.g., chemokines, matrix-released molecules, system challenges. Various pathophysiological conditions growth factors, etc). The response is either a migration Cold Spring Harbor (e.g., cancer, fibrosis, infections, chronic inflammation) usurp toward or away from environmental cues and this directional Conference -Neuronal and/or compromise the dynamic physiological processes response requires dynamic reorganization of the actin Circuts underlying migration1-5. cytoskeleton. A central question is the identity of the March 18-21st signaling molecule (or molecules) that relays the extracellular 1-4 Cold Spring Harbor, NY Rho-family GTPases (e.g., RhoA, Rac1, Cdc42, and RhoJ) act information to the actin cytoskeleton . -
Mutations in Desmin's Carboxy-Terminal “Tail” Domain Severely Modify Filament and Network Mechanics
doi:10.1016/j.jmb.2010.02.024 J. Mol. Biol. (2010) 397, 1188–1198 Available online at www.sciencedirect.com Mutations in Desmin's Carboxy-Terminal “Tail” Domain Severely Modify Filament and Network Mechanics Harald Bär1,2†, Michael Schopferer3†, Sarika Sharma1,2, Bernhard Hochstein3, Norbert Mücke4, Harald Herrmann2 and Norbert Willenbacher3⁎ 1Department of Cardiology, Inherited mutations in the gene coding for the intermediate filament protein University of Heidelberg, desmin have been demonstrated to cause severe skeletal and cardiac 69120 Heidelberg, Germany myopathies. Unexpectedly, some of the mutated desmins, in particular those carrying single amino acid alterations in the non-α-helical carboxy- 2Group Functional Architecture terminal domain (“tail”), have been demonstrated to form apparently of the Cell, German Cancer normal filaments both in vitro and in transfected cells. Thus, it is not clear if Research Center, filament properties are affected by these mutations at all. For this reason, we 69120 Heidelberg, Germany performed oscillatory shear experiments with six different desmin “tail” 3Institute of Mechanical Process mutants in order to characterize the mesh size of filament networks and Engineering and Mechanics, their strain stiffening properties. Moreover, we have carried out high- University of Karlsruhe, frequency oscillatory squeeze flow measurements to determine the bending Gotthard-Franz-Straße 3, 76131 stiffness of the respective filaments, characterized by the persistence length Karlsruhe, Germany lp. Interestingly, mesh size was not altered for the mutant filament 4 networks, except for the mutant DesR454W, which apparently did not Division of Biophysics of form proper filament networks. Also, the values for bending stiffness were Macromolecules, German “ ” – μ in the same range for both the tail mutants (lp =1.0 2.0 m) and the wild- Cancer Research Center, μ type desmin (lp =1.1±0.5 m). -
Structure and Mechanics of Proteins from Single Molecules to Cells
University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations Summer 2009 Structure and Mechanics of Proteins from Single Molecules to Cells Andre E. Brown University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Biological and Chemical Physics Commons Recommended Citation Brown, Andre E., "Structure and Mechanics of Proteins from Single Molecules to Cells" (2009). Publicly Accessible Penn Dissertations. 9. https://repository.upenn.edu/edissertations/9 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/9 For more information, please contact [email protected]. Structure and Mechanics of Proteins from Single Molecules to Cells Abstract Physical factors drive evolution and play important roles in motility and attachment as well as in differentiation. As animal cells adhere to survive, they generate force and “feel” various mechanical features of their surroundings and respond to externally applied forces. This mechanosensitivity requires a substrate for cells to adhere to and a mechanism for cells to apply force, followed by a cellular response to the mechanical properties of the substrate. We have taken an outside-in approach to characterize several aspects of cellular mechanosensitivity. First, we used single molecule force spectroscopy to measure how fibrinogen, an extracellular matrix protein that forms the scaffold of blood clots, responds to applied force and found that it rapidly unfolds in 23 nm steps at forces around 100 pN. Second, we used tensile testing to measure the force-extension behavior of fibrin gels and found that they behave almost linearly to strains of over 100%, have extensibilities of 170 ± 15 %, and undergo a large volume decrease that corresponds to a large and negative peak in compressibility at low strain, which indicates a structural transition. -
Soft Matter PAPER
View Article Online / Journal Homepage / Table of Contents for this issue Soft Matter Dynamic Article LinksC< Cite this: Soft Matter, 2012, 8, 7446 www.rsc.org/softmatter PAPER Growth of curved and helical bacterial cells Hongyuan Jiang and Sean X. Sun* Received 26th February 2012, Accepted 17th May 2012 DOI: 10.1039/c2sm25452b A combination of cell wall growth and cytoskeletal protein action gives rise to the observed bacterial cell shape. Aside from the common rod-like and spherical shapes, bacterial cells can also adopt curved or helical geometries. To understand how curvature in bacteria is developed or maintained, we examine how Caulobacter crescentus obtains its crescent-like shape. Caulobacter cells with or without the cytoskeletal bundle crescentin, an intermediate filament-like protein, exhibit two distinct growth modes, curvature maintenance that preserves the radius of curvature and curvature relaxation that straightens the cell (Fig. 1). Using a proposed mechanochemical model, we show that bending and twisting of the crescentin bundle can influence the stress distribution in the cell wall, and lead to the growth of curved cells. In contrast, after crescentin bundle is disrupted, originally curved cells will slowly relax towards a straight rod over time. The model is able to quantitatively capture experimentally observed curvature dynamics. Furthermore, we show that the shape anisotropy of the cross-section of a curved cell is never greater than 4%, even in the presence of crescentin. 1. Introduction forces applied by external constraints generate curved cells. Strikingly, the growth modes of the cell with or without cres- Bacterial cell walls are built through a complex biochemical centin are different17,18 as shown in Fig. -
A Metabolic Assembly Line in Bacteria
NEWS AND VIEWS A metabolic assembly line in bacteria Matthew T. Cabeen and Christine Jacobs-Wagner The bacterial cytoplasm is rich in filament-forming proteins, from homologues of eukaryotic cytoskeletal elements to other scaffolding and segregation proteins. We now learn that even the metabolic enzyme CTP synthase forms cytoplasmic filaments that affect bacterial cell shape. Bacteria keep surprising us. It was not so long in mediating cell curvature in Caulobacter cres- and analysing their function later. Using high- ago that they were thought to be mere bags of centus9; subsequent characterization revealed resolution electron cryotomography (ECT), an chemicals, possessing only the cell wall as a sort its intermediate filament-like properties9. But unbiased method which uses no labels, Jensen of exoskeleton to hold everything together. As what about proteins with functions that would and colleagues uncovered several filament-like it turns out, bacterial cells have a sophisticated never suggest any polymerizing property? structures in the cytoplasm of C. crescentus internal organization. They possess counter- Recent work has approached the discovery of that could not be identified by disrupting or parts of tubulin, actin and intermediate fila- subcellular structures from the opposite direc- eliminating known cytoskeletal structures10. ment proteins, suggesting that a cytoskeleton tion by searching for filamentous structures first Meanwhile, in another unbiased approach, first evolved in bacteria. Moreover, in recent years the known bacterial filament-forming proteins have expanded beyond the traditional cytoskeleton to include DNA segregators, structural scaffolds and proteins, the function of which are still unknown. On page 739 of this TubZ issue, Ingerson-Mahar et al. -
Neurofilaments: Neurobiological Foundations for Biomarker Applications
Neurofilaments: neurobiological foundations for biomarker applications Arie R. Gafson1, Nicolas R. Barthelmy2*, Pascale Bomont3*, Roxana O. Carare4*, Heather D. Durham5*, Jean-Pierre Julien6,7*, Jens Kuhle8*, David Leppert8*, Ralph A. Nixon9,10,11,12*, Roy Weller4*, Henrik Zetterberg13,14,15,16*, Paul M. Matthews1,17 1 Department of Brain Sciences, Imperial College, London, UK 2 Department of Neurology, Washington University School of Medicine, St Louis, MO, USA 3 a ATIP-Avenir team, INM, INSERM , Montpellier university , Montpellier , France. 4 Clinical Neurosciences, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, United Kingdom 5 Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Québec, Canada 6 Department of Psychiatry and Neuroscience, Laval University, Quebec, Canada. 7 CERVO Brain Research Center, 2601 Chemin de la Canardière, Québec, QC, G1J 2G3, Canada 8 Neurologic Clinic and Policlinic, Departments of Medicine, Biomedicine and Clinical Research, University Hospital Basel, University of Basel, Basel, Switzerland. 9 Center for Dementia Research, Nathan Kline Institute, Orangeburg, NY, 10962, USA. 10Departments of Psychiatry, New York University School of Medicine, New York, NY, 10016, 11 Neuroscience Institute, New York University School of Medicine, New York, NY, 10016, USA. 12Department of Cell Biology, New York University School of Medicine, New York, NY, 10016, USA 13 University College London Queen Square Institute of Neurology, London, UK 14 UK Dementia Research Institute at University College London 15 Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, the Sahlgrenska Academy at the University of Gothenburg, Mölndal, Sweden 16 Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, Mölndal, Sweden 17 UK Dementia Research Institute at Imperial College, London * Co-authors ordered alphabetically Address for correspondence: Prof. -
Cytoskeleton and Cell Motility
Cytoskeleton and Cell Motility Thomas Risler1 1Laboratoire Physicochimie Curie (CNRS-UMR 168), Institut Curie, Section de Recherche, 26 rue d’Ulm, 75248 Paris Cedex 05, France (Dated: January 23, 2008) PACS numbers: 1 Article Outline Glossary I. Definition of the Subject and Its Importance II. Introduction III. The Diversity of Cell Motility A. Swimming B. Crawling C. Extensions of cell motility IV. The Cell Cytoskeleton A. Biopolymers B. Molecular motors C. Motor families D. Other cytoskeleton-associated proteins E. Cell anchoring and regulatory pathways F. The prokaryotic cytoskeleton V. Filament-Driven Motility A. Microtubule growth and catastrophes B. Actin gels C. Modeling polymerization forces D. A model system for studying actin-based motility: The bacterium Listeria mono- cytogenes E. Another example of filament-driven amoeboid motility: The nematode sperm cell VI. Motor-Driven Motility A. Generic considerations 2 B. Phenomenological description close to thermodynamic equilibrium C. Hopping and transport models D. The two-state model E. Coupled motors and spontaneous oscillations F. Axonemal beating VII. Putting It Together: Active Polymer Solutions A. Mesoscopic approaches B. Microscopic approaches C. Macroscopic phenomenological approaches: The active gels D. Comparisons of the different approaches to describing active polymer solutions VIII. Extensions and Future Directions Acknowledgments Bibliography Glossary Cell Structural and functional elementary unit of all life forms. The cell is the smallest unit that can be characterized as living. Eukaryotic cell Cell that possesses a nucleus, a small membrane-bounded compartment that contains the genetic material of the cell. Cells that lack a nucleus are called prokaryotic cells or prokaryotes. Domains of life archaea, bacteria and eukarya - or in English eukaryotes, and made of eukaryotic cells - which constitute the three fundamental branches in which all life forms are classified. -
The Bacterial Actin Mreb Rotates, and Rotation Depends on Cell-Wall Assembly
The bacterial actin MreB rotates, and rotation depends on cell-wall assembly Sven van Teeffelena,1, Siyuan Wanga,b, Leon Furchtgottc,d, Kerwyn Casey Huangd, Ned S. Wingreena,b, Joshua W. Shaevitzb,e,1, and Zemer Gitaia,1 aDepartment of Molecular Biology, Princeton University, Princeton, NJ 08544; bLewis–Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544; cBiophysics Program, Harvard University, Cambridge, MA 02138; dDepartment of Bioengineering, Stanford University, Stanford, CA 94305; and eDepartment of Physics, Princeton University, Princeton, NJ 08854 Edited by Lucy Shapiro, Stanford University School of Medicine, Palo Alto, CA, and approved July 27, 2011 (received for review June 3, 2011) Bacterial cells possess multiple cytoskeletal proteins involved in a tently in a nearly circumferential direction. Interestingly, this wide range of cellular processes. These cytoskeletal proteins are MreB rotation is not driven by its own polymerization, but rather dynamic, but the driving forces and cellular functions of these requires cell-wall synthesis. These findings indicate that a motor dynamics remain poorly understood. Eukaryotic cytoskeletal dy- whose activity depends on cell-wall assembly rotates MreB. namics are often driven by motor proteins, but in bacteria no mo- Furthermore, the coupling of MreB rotation to cell-wall synthesis tors that drive cytoskeletal motion have been identified to date. suggests that MreB may not merely act upstream of cell-wall Here, we quantitatively study the dynamics of the Escherichia coli assembly. Indeed, computational simulations suggest that cou- actin homolog MreB, which is essential for the maintenance of pling MreB rotation to cell-wall synthesis can help cells maintain rod-like cell shape in bacteria.