| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print May 21, 2002
J. Biol. Chem, 10.1074/jbc.C200240200
Submitted on April 18, 2002
Revised on May 13, 2002
Accepted on May 21, 2002
12 directly binds to the SH3 domain and regulates paracellular permeability in epithelial cells
Renal Division, Brigham and Women's Hospital, Boston, MA 02115
Corresponding Author: tmeyer{at}ucsd.edu
Zona-Occludens proteins are multi-domain proteins usually localized at sites of intercellular junctions, yet little is known about their role in regulating junctional properties. Multiple signaling proteins regulate the junctional complex and several (including G proteins) have been co-localized with Zonula occludens-1 (ZO-1) in the tight junction of epithelial cells. However, evidence for direct interactions between signaling proteins and tight junction proteins has been lacking. In these studies, we constructed G
-GST fusion proteins and tested for interactions with [S35]-methionine labeled in-vitro translated ZO-1 and ZO-2. Only G12 directly interacted with in-vitro translated ZO-1 and ZO-2. Utilizing a series of ZO-1 domains expressed as GST fusion proteins and in-vitro translated [S35]-methionine labeled G12, we found that G12 and constitutively active (Q229L) 12 bind to the SH3 domain of ZO-1. This binding was not detected with SH3 domains from other proteins. Inducible expression of wildtype 12 and QL12 in MDCK cells was established using the Tet-Off system. In G12 expressing cells, we find that ZO-1 and G12 co-localize by confocal microscopy, co-immunoprecipitate, and that G12 from MDCK cell lysates binds to the GST-ZO-1-SH3 domain. Finally, we demonstrate that expression of QL12 reversibly increases paracellular permeability (decreased transepithelial resistance and increased paracellular flux) in MDCK cells. Consistent with ZO-1 as a scaffold for signaling molecules, these studies indicate that ZO-1 directly interacts with G12, and that G12 regulates barrier function of MDCK cells.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  E. Sabath, H. Negoro, S. Beaudry, M. Paniagua, S. Angelow, J. Shah, N. Grammatikakis, A. S. L. Yu, and B. M. Denker G{alpha}12 regulates protein interactions within the MDCK cell tight junction and inhibits tight-junction assembly J. Cell Sci., March 15, 2008; 121(6): 814 - 824. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Yanamadala, H. Negoro, L. Gunaratnam, T. Kong, and B. M. Denker G{alpha}12 Stimulates Apoptosis in Epithelial Cells through JNK1-mediated Bcl-2 Degradation and Up-regulation of I{kappa}B{alpha} J. Biol. Chem., August 17, 2007; 282(33): 24352 - 24363. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Zhu, R. I. Tate, R. Ruediger, T. E. Meigs, and B. M. Denker Domains Necessary for G{alpha}12 Binding and Stimulation of Protein Phosphatase-2A (PP2A): Is G{alpha}12 a Novel Regulatory Subunit of PP2A? Mol. Pharmacol., May 1, 2007; 71(5): 1268 - 1276. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Broughman, L. Sun, S. Umar, J. H. Sellin, and A. P. Morris Chronic PKC-beta2 activation in HT-29 Cl.19a colonocytes prevents cAMP-mediated ion secretion by inhibiting apical membrane CFTR targeting Am J Physiol Gastrointest Liver Physiol, August 1, 2006; 291(2): G331 - G344. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. McNeil, C. T. Capaldo, and I. G. Macara Zonula Occludens-1 Function in the Assembly of Tight Junctions in Madin-Darby Canine Kidney Epithelial Cells Mol. Biol. Cell, April 1, 2006; 17(4): 1922 - 1932. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Andreeva, R. Vaiskunaite, M. A. Kutuzov, J. Profirovic, R. A. Skidgel, and T. Voyno-Yasenetskaya Novel Mechanisms of G Protein-Dependent Regulation of Endothelial Nitric-Oxide Synthase Mol. Pharmacol., March 1, 2006; 69(3): 975 - 982. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. A. Kolosova, T. Mirzapoiazova, D. Adyshev, P. Usatyuk, L. H. Romer, J. R. Jacobson, V. Natarajan, D. B. Pearse, J. G.N. Garcia, and A. D. Verin Signaling Pathways Involved in Adenosine Triphosphate-Induced Endothelial Cell Barrier Enhancement Circ. Res., July 22, 2005; 97(2): 115 - 124. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. L. Hunt, M. R. Pollak, and B. M. Denker Cultured Podocytes Establish a Size-Selective Barrier Regulated by Specific Signaling Pathways and Demonstrate Synchronized Barrier Assembly in a Calcium Switch Model of Junction Formation J. Am. Soc. Nephrol., June 1, 2005; 16(6): 1593 - 1602. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. E. Meigs, J. Juneja, C. T. DeMarco, L. N. Stemmle, D. D. Kaplan, and P. J. Casey Selective Uncoupling of G{alpha}12 from Rho-mediated Signaling J. Biol. Chem., May 6, 2005; 280(18): 18049 - 18055. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Zhu, K. S. Kosik, T. E. Meigs, V. Yanamadala, and B. M. Denker G{alpha}12 Directly Interacts with PP2A: EVIDENCE FOR G{alpha}12-STIMULATED PP2A PHOSPHATASE ACTIVITY AND DEPHOSPHORYLATION OF MICROTUBULE-ASSOCIATED PROTEIN, Tau J. Biol. Chem., December 31, 2004; 279(53): 54983 - 54986. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. T. Kahle, G. G. MacGregor, F. H. Wilson, A. N. Van Hoek, D. Brown, T. Ardito, M. Kashgarian, G. Giebisch, S. C. Hebert, E. L. Boulpaep, et al. Paracellular Cl- permeability is regulated by WNK4 kinase: Insight into normal physiology and hypertension PNAS, October 12, 2004; 101(41): 14877 - 14882. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. D. Mruk and C. Y. Cheng Sertoli-Sertoli and Sertoli-Germ Cell Interactions and Their Significance in Germ Cell Movement in the Seminiferous Epithelium during Spermatogenesis Endocr. Rev., October 1, 2004; 25(5): 747 - 806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. E. Schneeberger and R. D. Lynch The tight junction: a multifunctional complex Am J Physiol Cell Physiol, June 1, 2004; 286(6): C1213 - C1228. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. N. Meyer, J. Hunt, C. Schwesinger, and B. M. Denker G{alpha}12 regulates epithelial cell junctions through Src tyrosine kinases Am J Physiol Cell Physiol, November 1, 2003; 285(5): C1281 - C1293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Waheed and T. L. Z. Jones Hsp90 Interactions and Acylation Target the G Protein Galpha 12 but Not Galpha 13 to Lipid Rafts J. Biol. Chem., August 30, 2002; 277(36): 32409 - 32412. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
