![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 10, Issue 12, 4177-4190, December 1999
and
*Medical Research Council-Laboratory for Molecular Cell Biology and
Department of Biochemistry and Molecular Biology, University College
London, London WC1E 6BT, United Kingdom; and Cell adhesion to individual macromolecules of the extracellular
matrix has dramatic effects on the subcellular localization of the
actin-bundling protein fascin and on the ability of cells to form
stable fascin microspikes. The actin-binding activity of fascin is
down-regulated by phosphorylation, and we used two differentiated cell
types, C2C12 skeletal myoblasts and LLC-PK1 kidney epithelial cells, to
examine the hypothesis that cell adhesion to the matrix components
fibronectin, laminin-1, and thrombospondin-1 differentially regulates
fascin phosphorylation. In both cell types, treatment with the PKC
activator 12-tetradecanoyl phorbol 13-acetate (TPA) or adhesion to
fibronectin led to a diffuse distribution of fascin after 1 h.
C2C12 cells contain the PKC family members Department
of Molecular Biology and Biochemistry, Rutgers University, Busch
Campus, Piscataway, New Jersey 08855
, 
, and 
, and
PKC localization was altered upon cell adhesion to fibronectin.
Two-dimensional isoelectric focusing/SDS-polyacrylamide gels were used
to determine that fascin became phosphorylated in cells adherent to
fibronectin and was inhibited by the PKC inhibitors calphostin C and
chelerythrine chloride. Phosphorylation of fascin was not detected in
cells adherent to thrombospondin-1 or to laminin-1. LLC-PK1 cells
expressing green fluorescent protein (GFP)-fascin also displayed
similar regulation of fascin phosphorylation. LLC-PK1 cells expressing
GFP-fascin S39A, a nonphosphorylatable mutant, did not undergo
spreading and focal contact organization on fibronectin, whereas cells
expressing a GFP-fascin S39D mutant with constitutive negative charge
spread more extensively than wild-type cells. In contrast, C2C12 cells
coexpressing S39A fascin with endogenous fascin remained competent to
form microspikes on thrombospondin-1, and cells that expressed fascin
S39D attached to thrombospondin-1 but did not form microspikes.
Blockade of PKC activity by TPA-induced down-regulation led to actin
association of wild-type fascin in fibronectin-adherent C2C12 and
LLC-PK1 cells but did not alter the distribution of S39A or S39D
fascins. The association of fascin with actin in fibronectin-adherent
cells was also evident in the presence of an inhibitory antibody to integrin 5 subunit. These novel results establish
matrix-initiated PKC-dependent regulation of fascin phosphorylation at
serine 39 as a mechanism whereby matrix adhesion is coupled to the
organization of cytoskeletal structure.
Corresponding author. E-mail address:
dmcbjca{at}ucl.ac.uk.
This article has been cited by other articles:
|
|
 |
|
  M. Parsons and J. C. Adams Rac regulates the interaction of fascin with protein kinase C in cell migration J. Cell Sci., September 1, 2008; 121(17): 2805 - 2813. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Hashimoto, M. Parsons, and J. C. Adams Dual Actin-bundling and Protein Kinase C-binding Activities of Fascin Regulate Carcinoma Cell Migration Downstream of Rac and Contribute to Metastasis Mol. Biol. Cell, November 1, 2007; 18(11): 4591 - 4602. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. S. Aratyn, T. E. Schaus, E. W. Taylor, and G. G. Borisy Intrinsic Dynamic Behavior of Fascin in Filopodia Mol. Biol. Cell, October 1, 2007; 18(10): 3928 - 3940. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Lin-Jones and B. Burnside Retina-Specific Protein Fascin 2 Is an Actin Cross-linker Associated with Actin Bundles in Photoreceptor Inner Segments and Calycal Processes Invest. Ophthalmol. Vis. Sci., March 1, 2007; 48(3): 1380 - 1388. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Vignjevic, S.-i. Kojima, Y. Aratyn, O. Danciu, T. Svitkina, and G. G. Borisy Role of fascin in filopodial protrusion J. Cell Biol., September 11, 2006; 174(6): 863 - 875. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Chakravarti, V. Sapountzi, and J. C. Adams Functional Role of Syndecan-1 Cytoplasmic V Region in Lamellipodial Spreading, Actin Bundling, and Cell Migration Mol. Biol. Cell, August 1, 2005; 16(8): 3678 - 3691. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-H. Disatnik, S. C. Boutet, W. Pacio, A. Y. Chan, L. B. Ross, C. H. Lee, and T. A. Rando The bi-directional translocation of MARCKS between membrane and cytosol regulates integrin-mediated muscle cell spreading J. Cell Sci., September 1, 2004; 117(19): 4469 - 4479. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. W. Zimmerman, J. M. D. T. Nelissen, S. E. van Emst-de Vries, P. H. G. M. Willems, F. de Lange, J. G. Collard, F. N. van Leeuwen, and C. G. Figdor Cytoskeletal restraints regulate homotypic ALCAM-mediated adhesion through PKC{alpha} independently of Rho-like GTPases J. Cell Sci., June 1, 2004; 117(13): 2841 - 2852. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. T. Kummer, T. Misgeld, J. W. Lichtman, and J. R. Sanes Nerve-independent formation of a topologically complex postsynaptic apparatus J. Cell Biol., March 29, 2004; 164(7): 1077 - 1087. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Loomis, L. Zheng, G. Sekerkova, B. Changyaleket, E. Mugnaini, and J. R. Bartles Espin cross-links cause the elongation of microvillus-type parallel actin bundles in vivo J. Cell Biol., December 8, 2003; 163(5): 1045 - 1055. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Vignjevic, D. Yarar, M. D. Welch, J. Peloquin, T. Svitkina, and G. G. Borisy Formation of filopodia-like bundles in vitro from a dendritic network J. Cell Biol., March 17, 2003; 160(6): 951 - 962. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Samstag, S. M. Eibert, M. Klemke, and G. H. Wabnitz Actin cytoskeletal dynamics in T lymphocyte activation and migration J. Leukoc. Biol., January 1, 2003; 73(1): 30 - 48. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Brandt, M. Gimona, M. Hillmann, H. Haller, and H. Mischak Protein Kinase C Induces Actin Reorganization via a Src- and Rho-dependent Pathway J. Biol. Chem., May 31, 2002; 277(23): 20903 - 20910. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-H. Disatnik, S. C. Boutet, C. H. Lee, D. Mochly-Rosen, and T. A. Rando Sequential activation of individual PKC isozymes in integrin-mediated muscle cell spreading: a role for MARCKS in an integrin signaling pathway J. Cell Sci., May 15, 2002; 115(10): 2151 - 2163. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Anilkumar, D. S. Annis, D. F. Mosher, and J. C. Adams Trimeric assembly of the C-terminal region of Thrombospondin-1 or Thrombospondin-2 is necessary for cell spreading and fascin spike organisation J. Cell Sci., January 6, 2002; 115(11): 2357 - 2366. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Masur, K. Lang, B. Niggemann, K. S. Zanker, and F. Entschladen High PKC {alpha} and Low E-Cadherin Expression Contribute to High Migratory Activity of Colon Carcinoma Cells Mol. Biol. Cell, July 1, 2001; 12(7): 1973 - 1982. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Adams, N. Kureishy, and A. L. Taylor A Role for Syndecan-1 in Coupling Fascin Spike Formation by Thrombospondin-1 J. Cell Biol., March 19, 2001; 152(6): 1169 - 1182. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Adams and M. A. Schwartz Stimulation of Fascin Spikes by Thrombospondin-1 Is Mediated by the GTPases Rac and Cdc42 J. Cell Biol., August 21, 2000; 150(4): 807 - 822. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Ku and K. E. Meier Phosphorylation of Paxillin via the ERK Mitogen-activated Protein Kinase Cascade in EL4 Thymoma Cells J. Biol. Chem., April 6, 2000; 275(15): 11333 - 11340. [Abstract] [Full Text] [PDF]
|
|
