Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Logo for

Mol. Biol. Cell 15 (12): 5538-5550

Copyright © 2004 by The American Society for Cell Biology.

Regulation of Epidermal Growth Factor Receptor Degradation by Heterotrimeric G{alpha}s Protein

Bin Zheng *, Christine Lavoie *, Ting-Dong Tang, Phuong Ma, Timo Meerloo, Anthony Beas, and Marilyn G. Farquhar {dagger}

Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0651

Received for publication June 3, 2004. Revision received September 15, 2004. Accepted for publication September 17, 2004.

Monitoring Editor: Juan S. Bonifacino

Abstract: Heterotrimeric G proteins have been implicated in the regulation of membrane trafficking, but the mechanisms involved are not well understood. Here, we report that overexpression of the stimulatory G protein subunit (G{alpha}s) promotes ligand-dependent degradation of epidermal growth factor (EGF) receptors and Texas Red EGF, and knock-down of G{alpha}s expression by RNA interference (RNAi) delays receptor degradation. We also show that G{alpha}s and its GTPase activating protein (GAP), RGS-PX1, interact with hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs), a critical component of the endosomal sorting machinery. G{alpha}s coimmunoprecipitates with Hrs and binds Hrs in pull-down assays. By immunofluorescence, exogenously expressed G{alpha}s colocalizes with myc-Hrs and GFP-RGS-PX1 on early endosomes, and expression of either Hrs or RGS-PX1 increases the localization of G{alpha}s on endosomes. Furthermore, knock-down of both Hrs and G{alpha}s by double RNAi causes greater inhibition of EGF receptor degradation than knock-down of either protein alone, suggesting that G{alpha}s and Hrs have cooperative effects on regulating EGF receptor degradation. These observations define a novel regulatory role for G{alpha}s in EGF receptor degradation and provide mechanistic insights into the function of G{alpha}s in endocytic sorting.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E04–06–0446. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E04–06–0446.

Abbreviations used: EGF, epidermal growth factor; EGFR, epidermal growth factor receptor; ESCRT, endosomal sorting complexes required for transport; GAP, GTPase activating protein; GPCR, G protein-coupled receptor; Hrs, hepatocyte growth factor-regulated tyrosine kinase substrate; MVB, multivesicular body; PFA, paraformaldehyde; PX, phoX domain; RGS, regulator of G protein signaling; RNAi, RNA interference; siRNA, small-interfering RNA; SNX, sorting nexin.

* These authors contributed equally to this work.

{dagger} Corresponding author. E-mail address: mfarquhar{at}ucsd.edu.

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
The Expanding Roles of G{beta}{gamma} Subunits in G Protein-Coupled Receptor Signaling and Drug Action.
S. M. Khan, R. Sleno, S. Gora, P. Zylbergold, J.-P. Laverdure, J.-C. Labbe, G. J. Miller, and T. E. Hebert (2013)
Pharmacol. Rev. 65, 545-577
   Abstract »    Full Text »    PDF »
G{alpha}s promotes EEA1 endosome maturation and shuts down proliferative signaling through interaction with GIV (Girdin).
A. O. Beas, V. Taupin, C. Teodorof, L. T. Nguyen, M. Garcia-Marcos, and M. G. Farquhar (2012)
Mol. Biol. Cell 23, 4623-4634
   Abstract »    Full Text »    PDF »
Peripherin Is a Subunit of Peripheral Nerve Neurofilaments: Implications for Differential Vulnerability of CNS and Peripheral Nervous System Axons.
A. Yuan, T. Sasaki, A. Kumar, C. M. Peterhoff, M. V. Rao, R. K. Liem, J.-P. Julien, and R. A. Nixon (2012)
J. Neurosci. 32, 8501-8508
   Abstract »    Full Text »    PDF »
Tyrosine Phosphorylation of the G{alpha}-Interacting Protein GIV Promotes Activation of Phosphoinositide 3-Kinase During Cell Migration.
C. Lin, J. Ear, Y. Pavlova, Y. Mittal, I. Kufareva, M. Ghassemian, R. Abagyan, M. Garcia-Marcos, and P. Ghosh (2011)
Science Signaling 4, ra64
   Abstract »    Full Text »    PDF »
Phox homology band 4.1/ezrin/radixin/moesin-like proteins function as molecular scaffolds that interact with cargo receptors and Ras GTPases.
R. Ghai, M. Mobli, S. J. Norwood, A. Bugarcic, R. D. Teasdale, G. F. King, and B. M. Collins (2011)
PNAS 108, 7763-7768
   Abstract »    Full Text »    PDF »
Characterization of PXK as a Protein Involved in Epidermal Growth Factor Receptor Trafficking.
H. Takeuchi, T. Takeuchi, J. Gao, L. C. Cantley, and M. Hirata (2010)
Mol. Cell. Biol. 30, 1689-1702
   Abstract »    Full Text »    PDF »
hNOA1 Interacts with Complex I and DAP3 and Regulates Mitochondrial Respiration and Apoptosis.
T. Tang, B. Zheng, S.-h. Chen, A. N. Murphy, K. Kudlicka, H. Zhou, and M. G. Farquhar (2009)
J. Biol. Chem. 284, 5414-5424
   Abstract »    Full Text »    PDF »
Activation of G{alpha}i3 triggers cell migration via regulation of GIV.
P. Ghosh, M. Garcia-Marcos, S. J. Bornheimer, and M. G. Farquhar (2008)
J. Cell Biol. 182, 381-393
   Abstract »    Full Text »    PDF »
Specific Leukotriene Receptors Couple to Distinct G Proteins to Effect Stimulation of Alveolar Macrophage Host Defense Functions.
C. M. Peres, D. M. Aronoff, C. H. Serezani, N. Flamand, L. H. Faccioli, and M. Peters-Golden (2007)
J. Immunol. 179, 5454-5461
   Abstract »    Full Text »    PDF »
Rin1 Interacts with Signal-transducing Adaptor Molecule (STAM) and Mediates Epidermal Growth Factor Receptor Trafficking and Degradation.
C. Kong, X. Su, P.-I Chen, and P. D. Stahl (2007)
J. Biol. Chem. 282, 15294-15301
   Abstract »    Full Text »    PDF »
Identification of Endosomal Epidermal Growth Factor Receptor Signaling Targets by Functional Organelle Proteomics.
T. Stasyk, N. Schiefermeier, S. Skvortsov, H. Zwierzina, J. Peranen, G. K. Bonn, and L. A. Huber (2007)
Mol. Cell. Proteomics 6, 908-922
   Abstract »    Full Text »    PDF »
Inactivation of neuregulin-1 by nitration.
D. E. Nethery, S. Ghosh, S. C. Erzurum, and J. A. Kern (2007)
Am J Physiol Lung Cell Mol Physiol 292, L287-L293
   Abstract »    Full Text »    PDF »
Hse1, a Component of the Yeast Hrs-STAM Ubiquitin-sorting Complex, Associates with Ubiquitin Peptidases and a Ligase to Control Sorting Efficiency into Multivesicular Bodies.
J. Ren, Y. Kee, J. M. Huibregtse, and R. C. Piper (2007)
Mol. Biol. Cell 18, 324-335
   Abstract »    Full Text »    PDF »
Essential role of RGS-PX1/sorting nexin 13 in mouse development and regulation of endocytosis dynamics.
B. Zheng, T. Tang, N. Tang, K. Kudlicka, K. Ohtsubo, P. Ma, J. D. Marth, M. G. Farquhar, and E. Lehtonen (2006)
PNAS 103, 16776-16781
   Abstract »    Full Text »    PDF »
Identification and Characterization of GIV, a Novel G{alpha}i/s -interacting Protein Found on COPI, Endoplasmic Reticulum-Golgi Transport Vesicles.
H. Le-Niculescu, I. Niesman, T. Fischer, L. DeVries, and M. G. Farquhar (2005)
J. Biol. Chem. 280, 22012-22020
   Abstract »    Full Text »    PDF »
{beta}-Adrenergic Receptor Stimulation Promotes G{alpha}s Internalization through Lipid Rafts: A Study in Living Cells.
J. A. Allen, J. Z. Yu, R. J. Donati, and M. M. Rasenick (2005)
Mol. Pharmacol. 67, 1493-1504
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882