| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print July 30, 2004
Cardiovascular Division, Brigham and Women's Hospital, Boston, MA 02115
Corresponding Author: tmichel{at}rics.bwh.harvard.edu
Caveolin-1 is a scaffolding/regulatory protein that interacts with diverse signaling molecules in endothelial cells. In order to explore the role of this protein in receptor-modulated signaling pathways, we transfected bovine aortic endothelial cells (BAEC) with small interfering RNA (siRNA) duplexes to down-regulate caveolin-1 expression. Transfection of BAEC with duplex siRNA targeted against caveolin-1 mRNA selectively “knocked-down” the expression of caveolin-1 by ~90%, as demonstrated by immunoblot analyses of BAEC lysates. We used discontinuous sucrose gradients to purify caveolin-containing lipid rafts from siRNA-treated endothelial cells. Despite the near-total down-regulation of caveolin-1 expression, the lipid raft targeting of diverse signaling proteins — including the endothelial isoform of nitric oxide synthase (eNOS), Src-family tyrosine kinases, Gaq and the insulin receptor — was unchanged. We explored the consequences of caveolin-1 knockdown on kinase pathways modulated by the agonists sphingosine-1 phosphate (S1P) and vascular endothelial growth factor (VEGF). siRNA-mediated caveolin-1 knockdown enhanced basal as well as S1P- and VEGF-induced phosphorylation of the protein kinase Akt, and did not modify the basal or agonist-induced phosphorylation of ERK1/2. Caveolin-1 knockdown also significantly enhanced the basal and agonist-induced activity of the small GTPase Rac. We used siRNA to down-regulate Rac expression in BAEC and we observed that Rac knockdown significantly reduced basal, S1P- and VEGF-induced Akt phosphorylation, suggesting a role for Rac activation in the caveolin siRNA-mediated increase in Akt phosphorylation. By using siRNA to knockdown caveolin-1 and Rac expression in cultured endothelial cells, we have found that caveolin-1 does not appear to be required for the targeting of signaling molecules to caveolae/lipid rafts and that caveolin-1 differentially modulates specific kinase pathways in endothelial cells.
J. Biol. Chem, 10.1074/jbc.M407051200
Submitted on June 23, 2004
Revised on July 28, 2004
Accepted on July 29, 2004
siRNA-mediated down-regulation of caveolin-1 differentially modulates signaling pathways in endothelial cells
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  L. C. Matthews, M. J. Taggart, and M. Westwood Modulation of Caveolin-1 Expression Can Affect Signalling through the Phosphatidylinositol 3-Kinase/Akt Pathway and Cellular Proliferation in Response to Insulin-Like Growth Factor I Endocrinology, October 1, 2008; 149(10): 5199 - 5208. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Cai, H. Wang, Y. Chen, L. Liu, W. T Gunning, L. E. M. Quintas, and Z.-J. Xie Regulation of caveolin-1 membrane trafficking by the Na/K-ATPase J. Cell Biol., September 22, 2008; 182(6): 1153 - 1169. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Chen, Y. C. Levine, D. E. Golan, T. Michel, and A. J. Lin Atrial Natriuretic Peptide-initiated cGMP Pathways Regulate Vasodilator-stimulated Phosphoprotein Phosphorylation and Angiogenesis in Vascular Endothelium J. Biol. Chem., February 15, 2008; 283(7): 4439 - 4447. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Salani, L. Briatore, S. Garibaldi, R. Cordera, and D. Maggi Caveolin-1 Down-Regulation Inhibits Insulin-Like Growth Factor-I Receptor Signal Transduction in H9C2 Rat Cardiomyoblasts Endocrinology, February 1, 2008; 149(2): 461 - 465. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Hu, R. D. Ye, M. C. Dinauer, A. B. Malik, and R. D. Minshall Neutrophil caveolin-1 expression contributes to mechanism of lung inflammation and injury Am J Physiol Lung Cell Mol Physiol, February 1, 2008; 294(2): L178 - L186. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Kou and T. Michel Epinephrine Regulation of the Endothelial Nitric-oxide Synthase: ROLES OF RAC1 AND beta3-ADRENERGIC RECEPTORS IN ENDOTHELIAL NO SIGNALING J. Biol. Chem., November 9, 2007; 282(45): 32719 - 32729. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. F. Pilch, R. P. Souto, L. Liu, M. P. Jedrychowski, E. A. Berg, C. E. Costello, and S. P. Gygi Cellular spelunking: exploring adipocyte caveolae J. Lipid Res., October 1, 2007; 48(10): 2103 - 2111. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Predescu, D. N. Predescu, and A. B. Malik Molecular determinants of endothelial transcytosis and their role in endothelial permeability Am J Physiol Lung Cell Mol Physiol, October 1, 2007; 293(4): L823 - L842. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Yamamoto, N. Shimizu, S. Obi, S. Kumagaya, Y. Taketani, A. Kamiya, and J. Ando Involvement of cell surface ATP synthase in flow-induced ATP release by vascular endothelial cells Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1646 - H1653. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. C. Levine, G. K. Li, and T. Michel Agonist-modulated Regulation of AMP-activated Protein Kinase (AMPK) in Endothelial Cells: EVIDENCE FOR AN AMPK -> Rac1 -> Akt -> ENDOTHELIAL NITRIC-OXIDE SYNTHASE PATHWAY J. Biol. Chem., July 13, 2007; 282(28): 20351 - 20364. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Carlile-Klusacek and V. Rizzo Endothelial cytoskeletal reorganization in response to PAR1 stimulation is mediated by membrane rafts but not caveolae Am J Physiol Heart Circ Physiol, July 1, 2007; 293(1): H366 - H375. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Grande-Garcia, A. Echarri, J. de Rooij, N. B. Alderson, C. M. Waterman-Storer, J. M. Valdivielso, and M. A. del Pozo Caveolin-1 regulates cell polarization and directional migration through Src kinase and Rho GTPases J. Cell Biol., May 21, 2007; 177(4): 683 - 694. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Hunter and G. F. Nixon Spatial Compartmentalization of Tumor Necrosis Factor (TNF) Receptor 1-dependent Signaling Pathways in Human Airway Smooth Muscle Cells: LIPID RAFTS ARE ESSENTIAL FOR TNF-{alpha}-MEDIATED ACTIVATION OF RhoA BUT DISPENSABLE FOR THE ACTIVATION OF THE NF-{kappa}B AND MAPK PATHWAYS J. Biol. Chem., November 10, 2006; 281(45): 34705 - 34715. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Ushio-Fukai and R. W. Alexander Caveolin-Dependent Angiotensin II Type 1 Receptor Signaling in Vascular Smooth Muscle Hypertension, November 1, 2006; 48(5): 797 - 803. [Full Text] [PDF]
|
|
|
|
|
|
E. Gonzalez, R. Kou, and T. Michel Rac1 Modulates Sphingosine 1-Phosphate-mediated Activation of Phosphoinositide 3-Kinase/Akt Signaling Pathways in Vascular Endothelial Cells J. Biol. Chem., February 10, 2006; 281(6): 3210 - 3216. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Miyawaki-Shimizu, D. Predescu, J. Shimizu, M. Broman, S. Predescu, and A. B. Malik siRNA-induced caveolin-1 knockdown in mice increases lung vascular permeability via the junctional pathway Am J Physiol Lung Cell Mol Physiol, February 1, 2006; 290(2): L405 - L413. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Mehta and A. B. Malik Signaling Mechanisms Regulating Endothelial Permeability Physiol Rev, January 1, 2006; 86(1): 279 - 367. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. A. Spurlin and D. C. Thurmond Syntaxin 4 Facilitates Biphasic Glucose-Stimulated Insulin Secretion from Pancreatic {beta}-Cells Mol. Endocrinol., January 1, 2006; 20(1): 183 - 193. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Mineo, A. K. Gormley, I. S. Yuhanna, S. Osborne-Lawrence, L. L. Gibson, L. Hahner, R. V. Shohet, S. Black, J. E. Salmon, D. Samols, et al. Fc{gamma}RIIB Mediates C-Reactive Protein Inhibition of Endothelial NO Synthase Circ. Res., November 25, 2005; 97(11): 1124 - 1131. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Sen-Banerjee, S. Mir, Z. Lin, A. Hamik, G. B. Atkins, H. Das, P. Banerjee, A. Kumar, and M. K. Jain Kruppel-Like Factor 2 as a Novel Mediator of Statin Effects in Endothelial Cells Circulation, August 2, 2005; 112(5): 720 - 726. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Nagai, T. Sato, N. Akimoto, A. Ito, and M. Sumida Isolation and Identification of Histone H3 Protein Enriched in Microvesicles Secreted from Cultured Sebocytes Endocrinology, June 1, 2005; 146(6): 2593 - 2601. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Erwin, A. J. Lin, D. E. Golan, and T. Michel Receptor-regulated Dynamic S-Nitrosylation of Endothelial Nitric-oxide Synthase in Vascular Endothelial Cells J. Biol. Chem., May 20, 2005; 280(20): 19888 - 19894. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. T. Dobrowsky, S. Rouen, and C. Yu Altered Neurotrophism in Diabetic Neuropathy: Spelunking the Caves of Peripheral Nerve J. Pharmacol. Exp. Ther., May 1, 2005; 313(2): 485 - 491. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-L. Wang, D. Ye, T. E. Peterson, S. Cao, V. H. Shah, Z. S. Katusic, G. C. Sieck, and H.-C. Lee Caveolae Targeting and Regulation of Large Conductance Ca2+-activated K+ Channels in Vascular Endothelial Cells J. Biol. Chem., March 25, 2005; 280(12): 11656 - 11664. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
