The fully linked HTML version of this article has now been published.
JCS ePress
online publication date 3 Feb 2004
doi: 10.1242/jcs.00906
Research Article
Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments
John E. Eriksson*,
Tao He,
Amy V. Trejo-Skalli,
Ann-Sofi Härmälä-Braskén,
Jukka Hellman,
Ying-Hao Chou,
and
Robert D. Goldman
* Author for correspondence (e-mail: john.eriksson{at}utu.fi)
Intermediate filaments (IFs) continuously exchange between a small, depolymerized fraction of IF protein and fully polymerized IFs. To elucidate the possible role of phosphorylation in regulating this equilibrium, we disrupted the exchange of phosphate groups by specific inhibition of dephosphorylation and by specific phosphorylation and site-directed mutagenesis of two of the major in vivo phosphorylation sites determined in this study. Inhibition of type-1 (PP1) and type-2A (PP2A) protein phosphatases in BHK-21 fibroblasts with calyculin-A, induced rapid vimentin phosphorylation in concert with disassembly of the IF polymers into soluble tetrameric vimentin oligomers. This oligomeric composition corresponded to the oligopeptides released by cAMP-dependent kinase (PKA) following in vitro phosphorylation. Characterization of the 32P-labeled vimentin phosphopeptides, demonstrated Ser-4, Ser-6, Ser-7, Ser-8, Ser-9, Ser-38, Ser-41, Ser-71, Ser-72, Ser-418, Ser-429, Thr-456, and Ser-457 as significant in vivo phosphorylation sites. A number of the interphase-specific high turnover sites were shown to be in vitro phosphorylation sites for PKA and protein kinase C (PKC). The effect of presence or absence of phosphate groups on individual subunits was followed in vivo by microinjecting PKA-phosphorylated (primarily S38 and S72) and mutant vimentin (S38:A, S72:A), respectively. The PKA-phosphorylated vimentin showed a clearly decelerated filament formation in vivo, whereas obstruction of phosphorylation at these sites by site-directed mutagenesis had no significant effect on the incorporation rates of subunits into assembled polymers. Taken together, our results suggest that elevated phosphorylation regulates IF assembly in vivo by changing the equilibrium constant of subunit exchange towards a higher off-rate.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
|
|
 
Y.-H. Chou, W.-L. Kuo, M. R. Rosner, W.-J. Tang, and R. D. Goldman
Structural changes in intermediate filament networks alter the activity of insulin-degrading enzyme
FASEB J,
November 1, 2009;
23(11):
3734 - 3742.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Chang, K. Barlan, Y.-H. Chou, B. Grin, M. Lakonishok, A. S. Serpinskaya, D. K. Shumaker, H. Herrmann, V. I. Gelfand, and R. D. Goldman
The dynamic properties of intermediate filaments during organelle transport
J. Cell Sci.,
August 15, 2009;
122(16):
2914 - 2923.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q.-F. Li, A. M. Spinelli, and D. D. Tang
Cdc42GAP, reactive oxygen species, and the vimentin network
Am J Physiol Cell Physiol,
August 1, 2009;
297(2):
C299 - C309.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Sivaramakrishnan, J. L. Schneider, A. Sitikov, R. D. Goldman, and K. M. Ridge
Shear Stress Induced Reorganization of the Keratin Intermediate Filament Network Requires Phosphorylation by Protein Kinase C {zeta}
Mol. Biol. Cell,
June 1, 2009;
20(11):
2755 - 2765.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Aziz, J. F. Hess, M. S. Budamagunta, P. G. FitzGerald, and J. C. Voss
Head and Rod 1 Interactions in Vimentin: IDENTIFICATION OF CONTACT SITES, STRUCTURE, AND CHANGES WITH PHOSPHORYLATION USING SITE-DIRECTED SPIN LABELING AND ELECTRON PARAMAGNETIC RESONANCE
J. Biol. Chem.,
March 13, 2009;
284(11):
7330 - 7338.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. C. Nery, J. Zeng, B. P. Niland, J. Hewett, J. Farley, D. Irimia, Y. Li, G. Wiche, A. Sonnenberg, and X. O. Breakefield
TorsinA binds the KASH domain of nesprins and participates in linkage between nuclear envelope and cytoskeleton
J. Cell Sci.,
October 15, 2008;
121(20):
3476 - 3486.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. A. Whipple, E. M. Balzer, E. H. Cho, M. A. Matrone, J. R. Yoon, and S. S. Martin
Vimentin Filaments Support Extension of Tubulin-Based Microtentacles in Detached Breast Tumor Cells
Cancer Res.,
July 15, 2008;
68(14):
5678 - 5688.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Tilleman, K. Van Steendam, T. Cantaert, F. De Keyser, D. Elewaut, and D. Deforce
Synovial detection and autoantibody reactivity of processed citrullinated isoforms of vimentin in inflammatory arthritides
Rheumatology,
May 1, 2008;
47(5):
597 - 604.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. D. Tang
Intermediate filaments in smooth muscle
Am J Physiol Cell Physiol,
April 1, 2008;
294(4):
C869 - C878.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Kouloumenta, M. Mavroidis, and Y. Capetanaki
Proper Perinuclear Localization of the TRIM-like Protein Myospryn Requires Its Binding Partner Desmin
J. Biol. Chem.,
November 30, 2007;
282(48):
35211 - 35221.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Csortos, I. Kolosova, and A. D. Verin
Regulation of vascular endothelial cell barrier function and cytoskeleton structure by protein phosphatases of the PPP family
Am J Physiol Lung Cell Mol Physiol,
October 1, 2007;
293(4):
L843 - L854.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Wang, Q.-F. Li, Y. Anfinogenova, and D. D. Tang
Dissociation of Crk-associated substrate from the vimentin network is regulated by p21-activated kinase on ACh activation of airway smooth muscle
Am J Physiol Lung Cell Mol Physiol,
January 1, 2007;
292(1):
L240 - L248.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q.-F. Li, A. M. Spinelli, R. Wang, Y. Anfinogenova, H. A. Singer, and D. D. Tang
Critical Role of Vimentin Phosphorylation at Ser-56 by p21-activated Kinase in Vimentin Cytoskeleton Signaling
J. Biol. Chem.,
November 10, 2006;
281(45):
34716 - 34724.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Zhou, M. Cadrin, H. Herrmann, C.-H. Chen, R. J. Chalkley, A. L. Burlingame, and M. B. Omary
Keratin 20 Serine 13 Phosphorylation Is a Stress and Intestinal Goblet Cell Marker
J. Biol. Chem.,
June 16, 2006;
281(24):
16453 - 16461.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Chang, Y. Shav-Tal, T. Trcek, R. H. Singer, and R. D. Goldman
Assembling an intermediate filament network by dynamic cotranslation
J. Cell Biol.,
February 27, 2006;
172(5):
747 - 758.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. D'Ambrosio, S. Arena, G. Fulcoli, M. H. Scheinfeld, D. Zhou, L. D'Adamio, and A. Scaloni
Hyperphosphorylation of JNK-interacting Protein 1, a Protein Associated with Alzheimer Disease
Mol. Cell. Proteomics,
January 1, 2006;
5(1):
97 - 113.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. A. Marganski, S. S. Gangopadhyay, H.-D. Je, C. Gallant, and K. G. Morgan
Targeting of a Novel Ca+2/Calmodulin-Dependent Protein Kinase II Is Essential for Extracellular Signal-Regulated Kinase-Mediated Signaling in Differentiated Smooth Muscle Cells
Circ. Res.,
September 16, 2005;
97(6):
541 - 549.
[Abstract]
[Full Text]
[PDF]
|
|
|
© The Company of Biologists Ltd 2004
