![]()
|
|
||||||||
J. Biol. Chem., Vol. 276, Issue 39, 36163-36167, September 28, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
, and
From the Department of Physiology, University of Tennessee Health
Science Center, Memphis, Tennessee 38163 and the
We have previously shown that tyrosine
phosphorylation of the actin-regulatory protein villin is accompanied
by the redistribution of phosphorylated villin and a concomitant
decrease in the F-actin content of intestinal epithelial cells. The
temporal and spatial correlation of these two events suggested that
tyrosine phosphorylation of villin may be involved in the rearrangement
of the microvillar cytoskeleton. This hypothesis was investigated by
analyzing the effects of tyrosine phosphorylation of villin on the
kinetics of actin polymerization by reconstituting in vitro
the tyrosine phosphorylation of villin and its association with actin.
Full-length recombinant human villin was phosphorylated in
vitro by expression in the TKX1-competent cells that carry an
inducible tyrosine kinase gene. The actin-binding properties
of villin were examined using a co-sedimentation assay. Phosphorylation
of villin did not change the stoichiometry (1:2) but decreased the
binding affinity (4.4 µM for unphosphorylated
versus 0.6 µM for phosphorylated) of villin for actin. Using a pyrene-actin-based fluorescence assay, we
demonstrated that tyrosine phosphorylation had a negative effect on
actin nucleation by villin. In contrast, tyrosine phosphorylation
enhanced actin severing by villin. Electron microscopic analysis showed
complementary morphological changes. Phosphorylation inhibited the
actin bundling and enhanced the actin severing functions of villin.
Taken together our data show that tyrosine phosphorylation of villin
decreases the amount of villin bound to actin filaments, inhibits the
actin-polymerizing properties of villin, and promotes the
actin-depolymerizing functions instead. These observations suggest a
role for tyrosine phosphorylation in modulating the microvillar
cytoskeleton in vivo by villin in response to specific
physiological stimuli.
Department of Biophysics and Biophysical Chemistry, Johns
Hopkins University, Baltimore, Maryland 21205
This article has been cited by other articles:
![]() |
S. P. George, Y. Wang, S. Mathew, K. Srinivasan, and S. Khurana Dimerization and Actin-bundling Properties of Villin and Its Role in the Assembly of Epithelial Cell Brush Borders J. Biol. Chem., September 7, 2007; 282(36): 26528 - 26541. [Abstract] [Full Text] [PDF] |
||||
![]() |
Y. Wang, A. Tomar, S. P. George, and S. Khurana Obligatory role for phospholipase C-{gamma}1 in villin-induced epithelial cell migration Am J Physiol Cell Physiol, May 1, 2007; 292(5): C1775 - C1786. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. Revenu, M. Courtois, A. Michelot, C. Sykes, D. Louvard, and S. Robine Villin Severing Activity Enhances Actin-based Motility In Vivo Mol. Biol. Cell, March 1, 2007; 18(3): 827 - 838. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. Tomar, S. George, P. Kansal, Y. Wang, and S. Khurana Interaction of Phospholipase C-{gamma}1 with Villin Regulates Epithelial Cell Migration J. Biol. Chem., October 20, 2006; 281(42): 31972 - 31986. [Abstract] [Full Text] [PDF] |
||||
![]() |
L. Heiska and O. Carpen Src Phosphorylates Ezrin at Tyrosine 477 and Induces a Phosphospecific Association between Ezrin and a Kelch-Repeat Protein Family Member J. Biol. Chem., March 18, 2005; 280(11): 10244 - 10252. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Gonzalez-Quevedo, M. Shoffer, L. Horng, and A. E. Oro Receptor tyrosine phosphatase-dependent cytoskeletal remodeling by the hedgehog-responsive gene MIM/BEG4 J. Cell Biol., January 31, 2005; 168(3): 453 - 463. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. Tomar, Y. Wang, N. Kumar, S. George, B. Ceacareanu, A. Hassid, K. E. Chapman, A. M. Aryal, C. M. Waters, and S. Khurana Regulation of Cell Motility by Tyrosine Phosphorylated Villin Mol. Biol. Cell, November 1, 2004; 15(11): 4807 - 4817. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. Kumar, A. Tomar, A. L. Parrill, and S. Khurana Functional Dissection and Molecular Characterization of Calcium-sensitive Actin-capping and Actin-depolymerizing Sites in Villin J. Biol. Chem., October 22, 2004; 279(43): 45036 - 45046. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. Kumar and S. Khurana Identification of a Functional Switch for Actin Severing by Cytoskeletal Proteins J. Biol. Chem., June 11, 2004; 279(24): 24915 - 24918. [Abstract] [Full Text] [PDF] |
||||
![]() |
W. Vermeulen, P. Vanhaesebrouck, M. Van Troys, M. Verschueren, F. Fant, M. Goethals, C. Ampe, J. C. Martins, and F. A.M. Borremans Solution structures of the C-terminal headpiece subdomains of human villin and advillin, evaluation of headpiece F-actin-binding requirements Protein Sci., May 1, 2004; 13(5): 1276 - 1287. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. Kumar, P. Zhao, A. Tomar, C. A. Galea, and S. Khurana Association of Villin with Phosphatidylinositol 4,5-Bisphosphate Regulates the Actin Cytoskeleton J. Biol. Chem., January 23, 2004; 279(4): 3096 - 3110. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Athman, D. Louvard, and S. Robine Villin Enhances Hepatocyte Growth Factor-induced Actin Cytoskeleton Remodeling in Epithelial Cells Mol. Biol. Cell, November 1, 2003; 14(11): 4641 - 4653. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. Nishizuka, S.-T. Chen, F. G. Gwadry, J. Alexander, S. M. Major, U. Scherf, W. C. Reinhold, M. Waltham, L. Charboneau, L. Young, et al. Diagnostic Markers That Distinguish Colon and Ovarian Adenocarcinomas: Identification by Genomic, Proteomic, and Tissue Array Profiling Cancer Res., September 1, 2003; 63(17): 5243 - 5250. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Athman, D. Louvard, and S. Robine The Epithelial Cell Cytoskeleton and Intracellular Trafficking: III. How is villin involved in the actin cytoskeleton dynamics in intestinal cells? Am J Physiol Gastrointest Liver Physiol, September 1, 2002; 283(3): G496 - G502. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. J. Kelso, A. M. Hudson, and L. Cooley Drosophila Kelch regulates actin organization via Src64-dependent tyrosine phosphorylation J. Cell Biol., February 18, 2002; 156(4): 703 - 713. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. J. Kelso, A. M. Hudson, and L. Cooley Drosophila Kelch regulates actin organization via Src64-dependent tyrosine phosphorylation J. Cell Biol., February 18, 2002; 156(4): 703 - 713. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |