Project

Back to overview

The role of amino-terminal sequences in actin isoform function

English title The role of amino-terminal sequences in actin isoform function
Applicant Chaponnier Christine
Number 125320
Funding scheme Project funding
Research institution Département de Pathologie et Immunologie Faculté de Médecine / CMU Université de Genève
Institution of higher education University of Geneva - GE
Main discipline Cellular Biology, Cytology
Start/End 01.04.2009 - 30.09.2013
Approved amount 427'386.00
Show all

All Disciplines (2)

Discipline
Cellular Biology, Cytology
Molecular Biology

Keywords (7)

Isoactin; Actin polymerization; Actin Binding Proteins; Myofibroblast; Fibrotic diseases; Tumor invasion; Cell migration and contraction

Lay Summary (English)

Lead
Lay summary
roles in nearly all aspects of eukaryotic cell biology. In vertebrates, six actin isoforms, i.e. two striated muscle (?-skeletal, ?-SKA and ?-cardiac, ?-CAA), two smooth muscle (?- and ?-SMA) and two cytoplasmic (?- and ?-actin) are encoded by distinct genes. All actin isoforms exhibit similar primary amino acid sequences and the main differences among them are located at the N-terminus. The major goal of our research is to elucidate the function of the actin isoforms during physiological activities and to examine whether their expression and/or structural organization are related to some pathological processes. We will investigate: 1) possible anti-fibrotic therapy by interfering on ?-SMA organization and expression in myofibroblasts. The myofibroblast, a differentiated fibroblast characterized by a high contractile apparatus and by the de novo expression of ?-SMA, is the master cell regulating the contraction phase during the wound healing process and fibrotic diseases. Novel formulation for the ?-SMA N-terminal peptide AcEEED cellular delivery will be tested. A library of approved FDA compounds will be screened and molecules will be selected for their inhibitory activity on ?-SMA expression and contractile activity; 2) In addition to its presence in vascular and enteric smooth muscle cells, ?-SMA expression was observed in a subset of myoepithelial cells in in situ breast cancer, whereas myofibroblasts in the stromal reaction or in intestinal fibrotic diseases were ?-SMA negative. We will analyze further the distribution ?-SMA in various myoepithelial cells, in particular in breast myoepithelial cells during development, lactation phases and tumor development phases; in enteric SMs in relation to intestinal obstructive diseases and intestinal cancers; 3) ?he regulation of ?- and ?-actin organization may be of major importance in tumor invasion processes: an increased ?-actin activity may play a leading role in the transition from an immotile to a motile cell; however, a reduced ?-actin activity may also contribute to facilitate the change to a more dynamic actin re-organization. In conclusion, our current results favor actin isoform functional diversities. The analysis of specific regulation of SMAs and cytoplasmic actins will likely assign specific roles to each isoactin in physiological and pathological situations.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Cooperation between β- and γ-cytoplasmic actins in the mechanical regulation of endothelial microparticle formation.
Latham Sharissa L, Chaponnier Christine, Dugina Vera, Couraud Pierre-Olivier, Grau Georges E R, Combes Valery (2013), Cooperation between β- and γ-cytoplasmic actins in the mechanical regulation of endothelial microparticle formation., in FASEB journal : official publication of the Federation of American Societies for Experimental Biolog, 27(2), 672-83.
Smooth muscle actin isoforms: a tug of war between contraction and compliance.
Arnoldi Richard, Hiltbrunner Anita, Dugina Vera, Tille Jean-Christophe, Chaponnier Christine (2013), Smooth muscle actin isoforms: a tug of war between contraction and compliance., in European journal of cell biology, 92(6-7), 187-200.
Smooth muscle hyperplasia due to loss of smooth muscle α-actin is driven by activation of focal adhesion kinase, altered p53 localization and increased levels of platelet-derived growth factor receptor-β.
Papke Christina L, Cao Jiumei, Kwartler Callie S, Villamizar Carlos, Byanova Katerina L, Lim Soon-Mi, Sreenivasappa Harini, Fischer Grant, Pham John, Rees Meredith, Wang Miranda, Chaponnier Christine, Gabbiani Giulio, Khakoo Aarif Y, Chandra Joya, Trache Andreea, Zimmer Warren, Milewicz Dianna M (2013), Smooth muscle hyperplasia due to loss of smooth muscle α-actin is driven by activation of focal adhesion kinase, altered p53 localization and increased levels of platelet-derived growth factor receptor-β., in Human molecular genetics, 22(15), 3123-37.
Actin isoforms and adhesion junctions reorganization in epithelial-mesenchymal transition of cervical carcinoma cells
Shagieva G., Domnina L., Chaponnier C., Dugina V. (2012), Actin isoforms and adhesion junctions reorganization in epithelial-mesenchymal transition of cervical carcinoma cells, in FEBS JOURNAL, 279, 317-317.
Actin isoforms and reorganization of adhesion junctions in epithelial-to-mesenchymal transition of cervical carcinoma cells
Shagieva G. S., Domnina L. V., Chipysheva T. A., Ermilova V. D., Chaponnier C., Dugina V. B. (2012), Actin isoforms and reorganization of adhesion junctions in epithelial-to-mesenchymal transition of cervical carcinoma cells, in BIOCHEMISTRY-MOSCOW, 77(11), 1266-1276.
Nonredundant roles of cytoplasmic beta- and gamma-actin isoforms in regulation of epithelial apical junctions
Baranwal Somesh, Naydenov Nayden G., Harris Gianni, Dugina Vera, Morgan Kathleen G., Chaponnier Christine, Ivanov Andrei I. (2012), Nonredundant roles of cytoplasmic beta- and gamma-actin isoforms in regulation of epithelial apical junctions, in MOLECULAR BIOLOGY OF THE CELL, 23(18), 3542-3553.
Resveratrol Treatment Reduces Cardiac Progenitor Cell Dysfunction and Prevents Morpho-Functional Ventricular Remodeling in Type-1 Diabetic Rats
Delucchi Francesca, Berni Roberta, Frati Caterina, Cavalli Stefano, Graiani Gallia, Sala Roberto, Chaponnier Christine, Gabbiani Giulio, Calani Luca, Del Rio Daniele, Bocchi Leonardo, Lagrasta Costanza, Quaini Federico, Stilli Donatella (2012), Resveratrol Treatment Reduces Cardiac Progenitor Cell Dysfunction and Prevents Morpho-Functional Ventricular Remodeling in Type-1 Diabetic Rats, in PLOS ONE, 7(6), 1-12.
Beta- and gamma-cytoplasmic actins are required for meiosis in mouse oocytes.
Brockmann Céline, Huarte Joachim, Dugina Vera, Challet Ludivine, Rey Emmanuelle, Conne Béatrice, Swetloff Adam, Nef Serge, Chaponnier Christine, Vassalli Jean-Dominique (2011), Beta- and gamma-cytoplasmic actins are required for meiosis in mouse oocytes., in Biology of reproduction, 85(5), 1025-39.
Sendai virus induced cytoplasmic actin remodeling correlates with efficient virus particle production.
Miazza Vincent, Mottet-Osman Geneviève, Startchick Sergei, Chaponnier Christine, Roux Laurent (2011), Sendai virus induced cytoplasmic actin remodeling correlates with efficient virus particle production., in Virology, 410(1), 7-16.
Tropomyosin variants describe distinct functional subcellular domains in differentiated vascular smooth muscle cells.
Gallant Cynthia, Appel Sarah, Graceffa Philip, Leavis Paul, Lin Jim Jung-Ching, Gunning Peter W, Schevzov Galina, Chaponnier Christine, DeGnore Jon, Lehman William, Morgan Kathleen G (2011), Tropomyosin variants describe distinct functional subcellular domains in differentiated vascular smooth muscle cells., in American journal of physiology. Cell physiology, 300(6), C1356-C136565.
γ-Actin regulates cell migration and modulates the ROCK signaling pathway.
Shum Michael S Y, Pasquier Eddy, Po'uha Sela T, O'Neill Geraldine M, Chaponnier Christine, Gunning Peter W, Kavallaris Maria (2011), γ-Actin regulates cell migration and modulates the ROCK signaling pathway., in FASEB journal : official publication of the Federation of American Societies for Experimental Biolog, 25(12), 4423-33.
Alpha actin isoforms expression in human and rat adult cardiac conduction system.
Orlandi Augusto, Hao Hiroyuki, Ferlosio Amedeo, Clément Sophie, Hirota Seiichi, Spagnoli Luigi Giusto, Gabbiani Giulio, Chaponnier Christine (2009), Alpha actin isoforms expression in human and rat adult cardiac conduction system., in Differentiation; research in biological diversity, 77(4), 360-8.
Beta and gamma-cytoplasmic actins display distinct distribution and functional diversity.
Dugina Vera, Zwaenepoel Ingrid, Gabbiani Giulio, Clément Sophie, Chaponnier Christine (2009), Beta and gamma-cytoplasmic actins display distinct distribution and functional diversity., in Journal of cell science, 122(Pt 16), 2980-8.
Immunohistochemical study of the phenotypic change of the mesenchymal cells during portal tract maturation in normal and fibrous (ductal plate malformation) fetal liver.
Villeneuve Julien, Pelluard-Nehme Fanny, Combe Chantal, Carles Dominique, Chaponnier Christine, Ripoche Jean, Balabaud Charles, Bioulac-Sage Paulette, Lepreux Sébastien (2009), Immunohistochemical study of the phenotypic change of the mesenchymal cells during portal tract maturation in normal and fibrous (ductal plate malformation) fetal liver., in Comparative hepatology, 8, 5-5.
Modulation of actin isoform expression before the transition from experimental compensated pressure-overload cardiac hypertrophy to decompensation.
Berni Roberta, Savi Monia, Bocchi Leonardo, Delucchi Francesca, Musso Ezio, Chaponnier Christine, Gabbiani Giulio, Clement Sophie, Stilli Donatella (2009), Modulation of actin isoform expression before the transition from experimental compensated pressure-overload cardiac hypertrophy to decompensation., in American journal of physiology. Heart and circulatory physiology, 296(5), H1625-H1632.
Re-expression of alpha skeletal actin as a marker for dedifferentiation in cardiac pathologies.
Driesen Ronald B, Verheyen Fons K, Debie Wiel, Blaauw Erik, Babiker Fawzi A, Cornelussen Richard N M, Ausma Jannie, Lenders Marie-Hélène, Borgers Marcel, Chaponnier Christine, Ramaekers Frans C S (2009), Re-expression of alpha skeletal actin as a marker for dedifferentiation in cardiac pathologies., in Journal of cellular and molecular medicine, 13(5), 896-908.

Collaboration

Group / person Country
Types of collaboration
Vascular Immunology Unit, Sydney Medical School, University of Sydney, Camperdown, New South Wales, Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Department of Microbiology and Molecular Medicine, University of Geneva, faculty of Medicine Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Department of Genetic Medicine and Development, University of Geneva, faculty of Medicine Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
A. N. Belozersky Institute of Physical and Chemical Biology, Moscow State University, Moscow, Russia Russia (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
European Cytoskeletal Forum, ECF2013 Poster 01.09.2013 Fribourg, Switzerland Chaponnier Christine;
European Cytoskeletal Forum ECF2013 Poster 01.09.2013 Fribourg, Switzerland Chaponnier Christine;
European Cytoskeletal forum, ECF2013 Poster 01.09.2013 Fribourg, Switzerland Hiltbrunner Anita; Chaponnier Christine; Arnoldi Richard;
Cytomeet, Berne Talk given at a conference 29.01.2013 Berne, Switzerland Arnoldi Richard; Chaponnier Christine;


Associated projects

Number Title Start Funding scheme
109879 The role of amino-terminal sequences in actin isoform function 01.10.2005 Project funding

Abstract

The major goal of our research is to elucidate the function of the actin isoforms during physiological activities and whether their expression and/or structural organization is related to some pathologies.During the last three years we have investigated: 1) the distribution and function of ß- and ?-cytoplasmic actins in fibroblastic and epithelial cells. Despite the fact that only 4 amino acid substitutions in position 1, 2, 3 and 9 differentiate these two isoforms, our results indicate, for the first time, that ß- and ?-cytoplasmic actins are polymerizing in different cellular structures. ß-cytoplasmic actin is detected at the basal at surface of the cells, in stress fibers and in circular bundles at the cell-cell contacts. On the other hand, ?-cytoplasmic actin is mainly organized under the dorsal membrane and as a dendritic network in motile cellular structures such as in lamellipodia. Isoform specific siRNA-mediated knockdown suggests that ß-cytoplasmic actin would play a preferential role in contractile activities whereas ?-cytoplasmic actin would mainly participate in the formation of a network necessary for cell shape flexibility and motile activity; 2) the distribution and function of a-smooth muscle actin (a-SMA) in cardiomyocytes. During myogenesis, knockdown of a-SMA in embryonic stem cells affects cardiac differentiation and the delivery of N-terminal a-SMA and a-skeletal actin peptides on beating embryoid bodies impairs frequency. a-SMA positive cardiomyocytes were also detected in the adult human and rat cardiac conduction system; 3) using a newly developed mAb, the distribution of ?-smooth muscle actin (?-SMA) in comparison to a-SMA expression. In addition to its presence in vascular and enteric smooth muscle cells, ?-SMA expression was observed in a subset of myoepithelial cells in in situ breast cancer, whereas myofibroblasts in the stromal reaction or in intestinal fibrotic diseases were ?-SMA negative.During the next three-year period, we plan to investigate: 1) possible anti-fibrotic therapies by interfering on a-SMA organization and expression in myofibroblasts. Novel formulation for the a-SMA N-terminal peptide AcEEED cellular delivery will be tested. A library of approved FDA compounds will be screened and molecules will be selected for their inhibitory activity on a-SMA expression and contractile activity; 2) ?-SMA distribution in various myoepithelial cells, in particular in breast myoepithelial cells during development, lactation phases and tumor development phases; in enteric SMs in relation to intestinal obstructive diseases and intestinal cancers; ?-SMA contractile activities in enteric smooth muscle cells in vitro in comparison to a-SMA. ?-SMA function using N-terminal peptide delivery and siRNA knockdown approaches; 3) ß- and ?-cytoplasmic actins distribution in different cell compartments using biochemical approaches; their regulation in more details during cytokinesis and cell attachment to substrate by confocal microscopy in relation to specific actin binding proteins.In conclusion, our current results favor actin isoform functional diversities. The analysis of specific regulation of SMAs and cytoplasmic actins will likely assign specific roles to each isoactin in physiological and pathological situations.
-