Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 333K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Hong-Geller, E. Articles by Cerione, R. A. Search for Related Content PubMed PubMed Citation Articles by Hong-Geller, E. Articles by Cerione, R. A. Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/2000//481 $5.00
The Journal of Cell Biology, Volume 148, Number 3, , 2000 481-494

Cdc42 and Rac Stimulate Exocytosis of Secretory Granules by Activating the Ip₃/Calcium Pathway in Rbl-2h3 Mast Cells

^a Department of Molecular Medicine, Cornell University, Ithaca, New York 14853
Veterinary Medical Center C3-155, Cornell University, Ithaca, NY 14853.(607) 253-3659(607) 253-3888

rac1{at}cornell.edu

We have expressed dominant-active and dominant-negative forms of the Rho GTPases, Cdc42 and Rac, using vaccinia virus to evaluate the effects of these mutants on the signaling pathway leading to the degranulation of secretory granules in RBL-2H3 cells. Dominant-active Cdc42 and Rac enhance antigen-stimulated secretion by about twofold, whereas the dominant-negative mutants significantly inhibit secretion. Interestingly, treatment with the calcium ionophore, A23187, and the PKC activator, PMA, rescues the inhibited levels of secretion in cells expressing the dominant-negative mutants, implying that Cdc42 and Rac act upstream of the calcium influx pathway. Furthermore, cells expressing the dominant-active mutants exhibit elevated levels of antigen-stimulated IP₃ production, an amplified antigen-stimulated calcium response consisting of both calcium release from internal stores and influx from the extracellular medium, and an increase in aggregate formation of the IP₃ receptor. In contrast, cells expressing the dominant-negative mutants display the opposite phenotypes. Finally, we are able to detect an in vitro interaction between Cdc42 and PLC1, the enzyme immediately upstream of IP₃ formation. Taken together, these findings implicate Cdc42 and Rac in regulating the exocytosis of secretory granules by stimulation of IP₃ formation and calcium mobilization upon antigen stimulation.

Key Words: Cdc42p • Rac • calcium signaling • degranulation • signal transduction

© 2000 The Rockefeller University Press

Abbreviations used in this paper: FAK, focal adhesion kinase; IP₃, inositol 1,4,5-trisphosphate; PBD, Cdc42/Rac (p21)-binding domain; PI3-K, phosphoinositide 3-kinase; PIP₂-4,5, phosphatidylinositol-4,5-bisphosphate; PKC, protein kinase C; PLC, phospholipase C-.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• Zhang, X.-F., Forscher, P. (2009). Rac1 Modulates Stimulus-evoked Ca2+ Release in Neuronal Growth Cones via Parallel Effects on Microtubule/Endoplasmic Reticulum Dynamics and Reactive Oxygen Species Production. Mol. Biol. Cell 20: 3700-3712 [Abstract] [Full Text]  
• Allen, J. D., Jaffer, Z. M., Park, S.-J., Burgin, S., Hofmann, C., Sells, M. A., Chen, S., Derr-Yellin, E., Michels, E. G., McDaniel, A., Bessler, W. K., Ingram, D. A., Atkinson, S. J., Travers, J. B., Chernoff, J., Clapp, D. W. (2009). p21-activated kinase regulates mast cell degranulation via effects on calcium mobilization and cytoskeletal dynamics. Blood 113: 2695-2705 [Abstract] [Full Text]  
• Momboisse, F., Lonchamp, E., Calco, V., Ceridono, M., Vitale, N., Bader, M.-F., Gasman, S. (2009). {beta}PIX-activated Rac1 stimulates the activation of phospholipase D, which is associated with exocytosis in neuroendocrine cells. J. Cell Sci. 122: 798-806 [Abstract] [Full Text]  
• Ferraro, F., Ma, X.-M., Sobota, J. A., Eipper, B. A., Mains, R. E. (2007). Kalirin/Trio Rho Guanine Nucleotide Exchange Factors Regulate a Novel Step in Secretory Granule Maturation. Mol. Biol. Cell 18: 4813-4825 [Abstract] [Full Text]  
• Cao, L., Yu, K., Banh, C., Nguyen, V., Ritz, A., Raphael, B. J., Kawakami, Y., Kawakami, T., Salomon, A. R. (2007). Quantitative Time-Resolved Phosphoproteomic Analysis of Mast Cell Signaling. J. Immunol. 179: 5864-5876 [Abstract] [Full Text]  
• El-Sibai, M., Nalbant, P., Pang, H., Flinn, R. J., Sarmiento, C., Macaluso, F., Cammer, M., Condeelis, J. S., Hahn, K. M., Backer, J. M. (2007). Cdc42 is required for EGF-stimulated protrusion and motility in MTLn3 carcinoma cells. J. Cell Sci. 120: 3465-3474 [Abstract] [Full Text]  
• Meyer, G. K. A., Neetz, A., Brandes, G., Tsikas, D., Butterfield, J. H., Just, I., Gerhard, R. (2007). Clostridium difficile Toxins A and B Directly Stimulate Human Mast Cells. Infect. Immun. 75: 3868-3876 [Abstract] [Full Text]  
• Pochynyuk, O., Staruschenko, A., Bugaj, V., LaGrange, L., Stockand, J. D. (2007). Quantifying RhoA Facilitated Trafficking of the Epithelial Na+ Channel toward the Plasma Membrane with Total Internal Reflection Fluorescence-Fluorescence Recovery after Photobleaching. J. Biol. Chem. 282: 14576-14585 [Abstract] [Full Text]  
• Pochynyuk, O., Medina, J., Gamper, N., Genth, H., Stockand, J. D., Staruschenko, A. (2006). Rapid Translocation and Insertion of the Epithelial Na+ Channel in Response to RhoA Signaling. J. Biol. Chem. 281: 26520-26527 [Abstract] [Full Text]  
• Mizota, K., Ueda, H. (2006). Endocrine Disrupting Chemical Atrazine Causes Degranulation through Gq/11 Protein-Coupled Neurosteroid Receptor in Mast Cells. Toxicol Sci 90: 362-368 [Abstract] [Full Text]  
• Suzuki, J., Jin, Z.-G., Meoli, D. F., Matoba, T., Berk, B. C. (2006). Cyclophilin A Is Secreted by a Vesicular Pathway in Vascular Smooth Muscle Cells. Circ. Res. 98: 811-817 [Abstract] [Full Text]  
• Alberts, P., Rudge, R., Irinopoulou, T., Danglot, L., Gauthier-Rouviere, C., Galli, T. (2006). Cdc42 and Actin Control Polarized Expression of TI-VAMP Vesicles to Neuronal Growth Cones and Their Fusion with the Plasma Membrane. Mol. Biol. Cell 17: 1194-1203 [Abstract] [Full Text]  
• Piechulek, T., Rehlen, T., Walliser, C., Vatter, P., Moepps, B., Gierschik, P. (2005). Isozyme-specific Stimulation of Phospholipase C-{gamma}2 by Rac GTPases. J. Biol. Chem. 280: 38923-38931 [Abstract] [Full Text]  
• Lominadze, G., Powell, D. W., Luerman, G. C., Link, A. J., Ward, R. A., McLeish, K. R. (2005). Proteomic Analysis of Human Neutrophil Granules. Mol. Cell. Proteomics 4: 1503-1521 [Abstract] [Full Text]  
• Gu, Y., Fu, Y., Dowd, P., Li, S., Vernoud, V., Gilroy, S., Yang, Z. (2005). A Rho family GTPase controls actin dynamics and tip growth via two counteracting downstream pathways in pollen tubes. JCB 169: 127-138 [Abstract] [Full Text]  
• Tateishi, Y., Hattori, M., Nakayama, T., Iwai, M., Bannai, H., Nakamura, T., Michikawa, T., Inoue, T., Mikoshiba, K. (2005). Cluster Formation of Inositol 1,4,5-Trisphosphate Receptor Requires Its Transition to Open State. J. Biol. Chem. 280: 6816-6822 [Abstract] [Full Text]  
• Nevins, A. K., Thurmond, D. C. (2005). A Direct Interaction between Cdc42 and Vesicle-associated Membrane Protein 2 Regulates SNARE-dependent Insulin Exocytosis. J. Biol. Chem. 280: 1944-1952 [Abstract] [Full Text]  
• Xin, X., Ferraro, F., Back, N., Eipper, B. A., Mains, R. E. (2004). Cdk5 and Trio modulate endocrine cell exocytosis. J. Cell Sci. 117: 4739-4748 [Abstract] [Full Text]  
• Abdel-Latif, D., Steward, M., Macdonald, D. L., Francis, G. A., Dinauer, M. C., Lacy, P. (2004). Rac2 is critical for neutrophil primary granule exocytosis. Blood 104: 832-839 [Abstract] [Full Text]  
• Bourguignon, L. Y. W., Singleton, P. A., Diedrich, F. (2004). Hyaluronan-CD44 Interaction with Rac1-dependent Protein Kinase N-{gamma} Promotes Phospholipase C{gamma}1 Activation, Ca2+ Signaling, and Cortactin-Cytoskeleton Function Leading to Keratinocyte Adhesion and Differentiation. J. Biol. Chem. 279: 29654-29669 [Abstract] [Full Text]  
• Li, J., O'Connor, K. L., Hellmich, M. R., Greeley, G. H. Jr., Townsend, C. M. Jr., Evers, B. M. (2004). The Role of Protein Kinase D in Neurotensin Secretion Mediated by Protein Kinase C-{alpha}/-{delta} and Rho/Rho Kinase. J. Biol. Chem. 279: 28466-28474 [Abstract] [Full Text]  
• Cho, S.-H., You, H.-J., Woo, C.-H., Yoo, Y.-J., Kim, J.-H. (2004). Rac and Protein Kinase C-{delta} Regulate ERKs and Cytosolic Phospholipase A2 in Fc{epsilon}RI Signaling to Cysteinyl Leukotriene Synthesis in Mast Cells. J. Immunol. 173: 624-631 [Abstract] [Full Text]  
• Chu, P. C., Wu, J., Liao, X. C., Pardo, J., Zhao, H., Li, C., Mendenhall, M. K., Pali, E., Shen, M., Yu, S., Taylor, V. C., Aversa, G., Molineaux, S., Payan, D. G., Masuda, E. S. (2004). A Novel Role for p21-Activated Protein Kinase 2 in T Cell Activation. J. Immunol. 172: 7324-7334 [Abstract] [Full Text]  
• Li, J., Luo, R., Kowluru, A., Li, G. (2004). Novel regulation by Rac1 of glucose- and forskolin-induced insulin secretion in INS-1 {beta}-cells. Am. J. Physiol. Endocrinol. Metab. 286: E818-E827 [Abstract] [Full Text]  
• Papakonstanti, E. A., Stournaras, C. (2004). Tumor Necrosis Factor-{alpha} Promotes Survival of Opossum Kidney Cells via Cdc42-induced Phospholipase C-{gamma}1 Activation and Actin Filament Redistribution. Mol. Biol. Cell 15: 1273-1286 [Abstract] [Full Text]  
• Li, Q., Ho, C. S, Marinescu, V., Bhatti, H., Bokoch, G. M, Ernst, S. A, Holz, R. W, Stuenkel, E. L (2003). Facilitation of Ca2+-dependent exocytosis by Rac1-GTPase in bovine chromaffin cells. J. Physiol. 550: 431-445 [Abstract] [Full Text]  
• Papakonstanti, E. A., Kampa, M., Castanas, E., Stournaras, C. (2003). A Rapid, Nongenomic, Signaling Pathway Regulates the Actin Reorganization Induced by Activation of Membrane Testosterone Receptors. Mol. Endocrinol. 17: 870-881 [Abstract] [Full Text]  
• Zhong, B., Jiang, K., Gilvary, D. L., Epling-Burnette, P. K., Ritchey, C., Liu, J., Jackson, R. J., Hong-Geller, E., Wei, S. (2003). Human neutrophils utilize a Rac/Cdc42-dependent MAPK pathway to direct intracellular granule mobilization toward ingested microbial pathogens. Blood 101: 3240-3248 [Abstract] [Full Text]  
• Djouder, N., Aneiros, E., Cavalie, A., Aktories, K. (2003). Effects of Large Clostridial Cytotoxins on Activation of RBL 2H3-hm1 Mast Cells Indicate Common and Different Roles of Rac in Fcepsilon RI and M1-Receptor Signaling. J. Pharmacol. Exp. Ther. 304: 1243-1250 [Abstract] [Full Text]  
• Camacho, L., Malho, R. (2003). Endo/exocytosis in the pollen tube apex is differentially regulated by Ca2+ and GTPases. J Exp Bot 54: 83-92 [Abstract] [Full Text]  
• Jiang, K., Zhong, B., Ritchey, C., Gilvary, D. L., Hong-Geller, E., Wei, S., Djeu, J. Y. (2003). Regulation of Akt-dependent cell survival by Syk and Rac. Blood 101: 236-244 [Abstract] [Full Text]  
• Hobert, M. E., Sands, K. A., Mrsny, R. J., Madara, J. L. (2002). Cdc42 and Rac1 Regulate Late Events in Salmonella typhimurium-induced Interleukin-8 Secretion from Polarized Epithelial Cells. J. Biol. Chem. 277: 51025-51032 [Abstract] [Full Text]  
• Humeau, Y., Popoff, M. R., Kojima, H., Doussau, F., Poulain, B. (2002). Rac GTPase Plays an Essential Role in Exocytosis by Controlling the Fusion Competence of Release Sites. J. Neurosci. 22: 7968-7981 [Abstract] [Full Text]  
• Rabiet, M.-J., Tardif, M., Braun, L., Boulay, F. (2002). Inhibitory effects of a dominant-interfering form of the Rho-GTPase Cdc42 in the chemoattractant-elicited signaling pathways leading to NADPH oxidase activation in differentiated HL-60 cells. Blood 100: 1835-1844 [Abstract] [Full Text]  
• Scott, G., Leopardi, S., Printup, S., Madden, B. C. (2002). Filopodia are conduits for melanosome transfer to keratinocytes. J. Cell Sci. 115: 1441-1451 [Abstract] [Full Text]  
• Vazquez-Martinez, R., Shorte, S. L., Faught, W. J., Leaumont, D. C., Frawley, L. S., Boockfor, F. R. (2001). Pulsatile Exocytosis Is Functionally Associated with GnRH Gene Expression in Immortalized GnRH-Expressing Cells. Endocrinology 142: 5364-5370 [Abstract] [Full Text]  
• Rojas, R., Ruiz, W. G., Leung, S.-M., Jou, T.-S., Apodaca, G. (2001). Cdc42-dependent Modulation of Tight Junctions and Membrane Protein Traffic in Polarized Madin-Darby Canine Kidney Cells. Mol. Biol. Cell 12: 2257-2274 [Abstract] [Full Text]  
• Manetz, T. S., Gonzalez-Espinosa, C., Arudchandran, R., Xirasagar, S., Tybulewicz, V., Rivera, J. (2001). Vav1 Regulates Phospholipase C{gamma} Activation and Calcium Responses in Mast Cells. Mol. Cell. Biol. 21: 3763-3774 [Abstract] [Full Text]  
• Djouder, N., Schmidt, G., Frings, M., Cavalie, A., Thelen, M., Aktories, K. (2001). Rac and Phosphatidylinositol 3-Kinase Regulate the Protein Kinase B in Fc{{epsilon}}RI Signaling in RBL 2H3 Mast Cells. J. Immunol. 166: 1627-1634 [Abstract] [Full Text]  
• Hong-Geller, E., Holowka, D., Siraganian, R. P., Baird, B., Cerione, R. A. (2001). Activated Cdc42/Rac reconstitutes Fcvarepsilon RI-mediated Ca2+ mobilization and degranulation in mutant RBL mast cells. Proc. Natl. Acad. Sci. USA 98: 1154-1159 [Abstract] [Full Text]  
• Field, K. A., Apgar, J. R., Hong-Geller, E., Siraganian, R. P., Baird, B., Holowka, D. (2000). Mutant RBL Mast Cells Defective in Fcepsilon RI Signaling and Lipid Raft Biosynthesis Are Reconstituted by Activated Rho-family GTPases. Mol. Biol. Cell 11: 3661-3673 [Abstract] [Full Text]  
• Klussmann, E., Tamma, G., Lorenz, D., Wiesner, B., Maric, K., Hofmann, F., Aktories, K., Valenti, G., Rosenthal, W. (2001). An Inhibitory Role of Rho in the Vasopressin-mediated Translocation of Aquaporin-2 into Cell Membranes of Renal Principal Cells. J. Biol. Chem. 276: 20451-20457 [Abstract] [Full Text]  
• Yang, W., Lo, C. G., Dispenza, T., Cerione, R. A. (2001). The Cdc42 Target ACK2 Directly Interacts with Clathrin and Influences Clathrin Assembly. J. Biol. Chem. 276: 17468-17473 [Abstract] [Full Text]  
• Powner, D. J., Hodgkin, M. N., Wakelam, M. J.O. (2002). Antigen-stimulated Activation of Phospholipase D1b by Rac1, ARF6, and PKCalpha in RBL-2H3 Cells. Mol. Biol. Cell 13: 1252-1262 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents