Rab1-Dependent Gβ1γ2 Trafficking during Muscle Mobilization


Fyn kinase-dependent cellular events were related with skeletal muscle denervation. In the present study, we used a combination of techniques to measure ER stability and the related Gβ1γ2 trafficking following muscle mobilization, and demonstrated a temporally and Fyn-dependent up-regulation of Ca2+ level in the mobilized muscle. In parallel, Fyn activity in ER was gradually decreased, which was accompanied by enhanced PTP1B activity and expressions of ER proteins (calnexin, Grp94, cyclophilin, and Hsp70). Moreover, during muscle mobilization, there was more membrane protein breakdown than protein synthesis, which probably featured robust Gβ1γ2 internalization, and Rab1-dependent transport into ER compartment; the signaling was related to disruption of PI3K-Akt signaling and decrement of muscular functions. Then, Gβ1γ2 trafficking is a key component necessary for the early recovery processes regarding muscle atrophy, which would be the therapeutic consideration for muscle repair and regeneration.

Share and Cite:

Huang, H. , Li, T. , Geng, L. and Zhao, H. (2015) Rab1-Dependent Gβ1γ2 Trafficking during Muscle Mobilization. Journal of Biomedical Science and Engineering, 8, 257-273. doi: 10.4236/jbise.2015.84025.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Fitts, R.H., Romatowski, J.G., Blaser, C., De La Cruz, L., Gettelman, G.J. and Widrick, J.J. (2000) Effect of Spaceflight on the Isotonic Contractile Properties of Single Skeletal Muscle Fibers in the Rhesus Monkey. Journal of Gravitational Physiology, 7, S53-S54.
[2] Marshall, J.M. and Tandon, H.C. (1984) Direct Observations of Muscle Arterioles and Venules Following Contraction of Skeletal Muscle Fibres in the Rat. Journal of Physiology, 350, 447-459.
[3] Bruns, A.F., Bao, L., Walker, J.H. and Ponnambalam, S. (2009) EGF-A-Stimulated Signalling in Endothelial Cells via a Dual Receptor Tyrosine Kinase System Is Dependent on Co-Ordinated Trafficking and Proteolysis. Biochemical Society Transactions, 37, 1193-1197.
[4] Prelle, K., Wobus, A.M., Krebs, O., Blum, W.F. and Wolf, E. (2000) Overexpression of Insulin-Like Growth Factor-II in Mouse Embryonic Stem Cells Promotes Myogenic Differentiation. Biochemical and Biophysical Research Communications, 277, 631-638.
[5] Rosen, K.M., Wentworth, B.M., Rosenthal, N. and Vil-la-Komaroff, L. (1993) Specific, Temporally Regulated Expression of the Insulin-Like Growth Factor II Gene During Muscle Cell Differentiation. Endocrinology, 133, 474-481.
[6] Hirashima, M., Ogawa, M., Nishikawa, S., Matsumura, K., Kawasaki, K., Shibuya, M. and Nishikawa, S. (2003) A Chemically Defined Culture of VEGFR2+ Cells Derived from Embryonic Stem Cells Reveals the Role of VEGFR1 in Tuning the Threshold for VEGF in Developing Endothelial Cells. Blood, 101, 2261-2267.
[7] Nielsena, S. and Pedersena, B.K. (2008) Skeletal Muscle as an Immunogenic Organ. Current Opinion in Pharmacology, 8, 346-351.
[8] Hanwei, H. and Zhao, H. (2010) FYN-Dependent Muscle-Immune Interaction after Sciatic Nerve Injury. Muscle & Nerve, 42, 70-77.
[9] Zhao, H. and Huang, H. (2012) Functional Capability of IL-15-Akt Signaling in the Denervated Muscle. Cytokine, 60, 608-615.
[10] Zhao, H., Huang, H.W., Wu, J.G. and Huang, P.Y. (2013) Specialization of Mitochondrial and Vascular Oxidant Modulated VEGFR in the Denervated Skeletal Muscle. Cell Signal, 25, 2106-2114.
[11] McGarrigle, D. and Huang, X.Y. (2007) GPCRs Signaling Directly through Src-Family Kinases. Science Signaling, 2007, pe35. http://dx.doi.org/10.1126/stke.3922007pe35
[12] Zhao, H., Wu, G. and Cao, X. (2013) EGFR Dependent Subcellular Communication Was Responsible for Morphine Mediated AC Superactivation. Cellular Signalling, 25, 417-428.
[13] Slessareva, J.E., Routt, S.M., Temple, B., Bankaitis, V.A. and Dohlman, H.G. (2006) Activation of the Phosphatidylinositol 3-Kinase Vps34 by a G Protein Alpha Subunit at the Endosome. Cell, 126, 191-203.
[14] Hunyady, L., Baukal, A.J., Gaborik, Z., Olivares-Reyes, J.A., Bor, M., Szaszak, M., Lodge, R., Catt, K.J. and Balla, T. (2002) Differential PI 3-Kinase Dependence of Early and Late Phases of Recycling of the Internalized AT1 Angiotensin Receptor. Journal of Cell Biology, 157, 1211-1222.
[15] Houle, S. and Marceau, F. (2003) Wortmannin Alters the Intracellular Trafficking of the Bradykinin B2 Receptor: Role of Phosphoinositide 3-Kinase and Rab5. Biochemical Journal, 375, 151-158.
[16] Kalia, M., Kumari, S., Chadda, R., Hill, M.M., Parton, R.G. and Mayor, S. (2006) Arf6-Independent GPI-Anchored Protein-Enriched Early Endosomal Compartments Fuse with Sorting Endosomes via a Rab5/phosphatidy-linositol-3'- kinase-Dependent Machinery. Molecular Biology of the Cell, 17, 3689-3704.
[17] Riek, R.P., Handschumacher, M.D., Sung, S.S., Tan, M., Glynias, M.J., Schluchter, M.D., Novotny, J. and Graham, R.M. (1995) Evolutionary Conservation of both the Hydrophilic and Hydrophobic Nature of Transmembrane Residues. Journal of Theoretical Biology, 172, 245-258.
[18] Hynes, T.R., Tang, L., Mervine, S.M., Sabo, J.L., Yost, E.A., Devreotes, P.N. and Berlot, C.H. (2004) Visualization of G Protein βγ Dimers Using Bimolecular Fluorescence Complementation Demonstrates Roles for Both β and γ in Subcellular Targeting. Journal of Biological Chemistry, 279, 30279-30286.
[19] Hynes, T.R., Mervine, S.M., Yost, E.A., Sabo, J.L. and Berlot, C.H. (2004) Live Cell Imaging of Gs and the β2-Adrenergic Receptor Demonstrates that Both Alphas and β1γ7 Internalize upon Stimulation and Exhibit Similar Trafficking Patterns That Differ from that of the β2-Adrenergic Receptor. Journal of Biological Chemistry, 279, 44101-44112.
[20] Saini, D.K., Kalyanaraman, V., Chisari, M. and Gautam, N. (2007) A Family of G Protein βγ Subunits Translocate Reversibly from the Plasma Membrane to Endomembranes on Receptor Activation. Journal of Biological Chemistry, 282, 24099-24108.
[21] Jamora, C., Yamanouye, N., Van Lint, J., Laudenslager, J., Vandenheede, J.R., Faulkner, D.J. and Malhotra, V. (1999) Gβγ-Mediated Regulation of Golgi Organization Is through the Direct Activation of Protein Kinase D. Cell, 98, 59-68. http://dx.doi.org/10.1016/S0092-8674(00)80606-6
[22] Díaz Anel, A.M. and Malhotra, V. (2005) PKCη Is Required for β1γ2/β3γ2- and PKD-Mediated Transport to the Cell Surface and the Organization of the Golgi apparatus. Journal of Cell Biology, 169, 83-91.
[23] Sonnichsen, B., De Renzis, S., Nielsen, E., Rietdorf, J. and Zerial, M. (2000) Distinct Membrane Domains on Endosomes in the Recycling Pathway Visualized by Multicolor Imaging of Rab4, Rab5, and Rab11. Journal of Cell Biology, 149, 901-914.
[24] Zerial, M. and McBride, H. (2001) Rab Proteins as Membrane Organizers. Nature Reviews Molecular Cell Biology, 2, 107-117.
[25] Miaczynska, M. and Zerial, M. (2002) Mosaic Organization of the Endocytic Pathway. Experimental Cell Research, 272, 8-14.
[26] Deldicque, L. (2013) Endoplasmic Reticulum Stress in Human Skeletal Muscle: Any Contribution to Sarcopenia? Frontiers in Physiology, 4, 236.
[27] Veeranki, S. and Tyagi, S.C. (2013) Defective Homocysteine Metabolism: Potential Implications for Skeletal Muscle Malfunction. International Journal of Molecular Sciences, 14, 15074-15091.
[28] Anderie, I., Schulz, I. and Schmid, A. (2007) Direct Interaction between ER Membrane-Bound PTP1B and Its Plasma Membrane-Anchored Targets. Cellular Signalling, 19, 582-592.
[29] Frangioni, J.V., Beahm, P.H., Shifrin, V., Jost, C.A. and Neel, B.G. (1992) The Nontransmembrane Tyrosine Phosphatase PTP-1B Localizes to the Endoplasmic Reticulum via Its 35 Amino Acid C-Terminal Sequence. Cell, 68, 545-560. http://dx.doi.org/10.1016/0092-8674(92)90190-N
[30] Frangioni, J.V., Oda, A., Smith, M., Salzman, E.W. and Neel, B.G. (1993) Calpain-Catalyzed Cleavage and Subcellular Relocation of Protein Phosphotyrosine Phosphatase 1B (PTP-1B) in Human Platelets. EMBO Journal, 12, 4843-4856.
[31] Haj, F.G., Verveer, P.J., Squire, A., Neel, B.G. and Bastiaens, P.I. (2002) Imaging Sites of Receptor Dephosphorylation by PTP1B on the Surface of the Endoplasmic Reticulum. Science, 295, 1708-1711.
[32] Hernandez, M.V., Sala, M.G., Balsamo, J., Lilien, J. and Arregui, C.O. (2006) ER-Bound PTP1B Is Targeted to Newly Forming Cell-Matrix Adhesions. Journal of Cell Science, 119, 1233-1243.
[33] Lorenzen, J.A., Dadabay, C.Y. and Fischer, E.H. (1995) COOH-Terminal Sequence Motifs Target the T Cell Protein Tyrosine Phosphatase to the ER and Nucleus. Journal of Cell Biology, 131, 631-643.
[34] Li, S., Depetris, R.S., Barford, D., Chernoff, J. and Hubbard, S.R. (2005) Crystal Structure of a Complex between Protein Tyrosine Phosphatase 1B and the Insulin Receptor Tyrosine Kinase. Structure, 13, 1643-1651.
[35] Tonks, N.K. (2003) PTP1B: From the Sidelines to the Front Lines! FEBS Letters, 546, 140-148.
[36] Gambardella, L. and Vermeren, S. (2013) Molecular Players in Neutrophil Chemotaxis—Focus on PI3K and Small GTPases. Journal of Leukocyte Biology, 94, 603-612.
[37] Hanna, S. and El-Sibai, M. (2013) Signaling Networks of Rho GTPases in Cell Motility. Cellular Signalling, 25, 1955-1961.
[38] Chua, C.E. and Tang, B.L. (2013) Linking Membrane Dynamics and Trafficking to Autophagy and the Unfolded Protein Response. Journal of Cellular Physiology, 228, 1638-1640.
[39] Liu, S. and Storrie, B. (2012) Are Rab Proteins the Link between Golgi Organization and Membrane Trafficking? Cellular and Molecular Life Sciences, 69, 4093-4106.
[40] Martinez, O. and Goud, B. (1998) Rab Proteins. Biochimica et Biophysica Acta, 1404, 101-112.
[41] Goud, B. and McCaffrey, M. (1991) Small GTP-Binding Proteins and Their Role in Transport. Current Opinion in Cell Biology, 3, 626-633.
[42] Punga, A.R., Maj, M., Lin, S., Meinen, S. and Ruegg, M.A. (2011) MuSK Levels Differ between Adult Skeletal Muscles and Influence Postsynaptic Plasticity. European Journal of Neuroscience, 33, 890-898.
[43] Rivas-Plata, K.A., Kraas, J.R., Saleh, S.M. and Swope, S.L. (2001) Src-Class Kinases Act within the Agrin/MuSK Pathway to Regulate Acetylcholine Receptor Phosphorylation, Cytoskeletal Anchoring, and Clustering. Journal of Neuroscience, 21, 3806-3818.
[44] Smith, C.L., Mittaud, P., Prescott, E.D., Fuhrer, C. and Burden, S.J. (2001) Src, Fyn, and Yes Are Not Required for Neuromuscular Synapse Formation but Are Necessary for Stabilization of Agrin-Induced Clusters of Acetylcholine Receptors. Journal of Neuroscience, 21, 3151-3160.
[45] Wang, Q., Zhang, B., Wang, Y.E., Xiong, W.C. and Mei, L. (2008) The Ig1/2 Domain of MuSK Binds to Muscle Surface and Is Involved in Acetylcholine Receptor Clustering. Neurosignals, 16, 246-253.
[46] Willmann, R., Pun, S., Stallmach, L., Sadasivam, G., Santos, A.F., Caroni, P. and Fuhrer, C. (2006) Cholesterol and Lipid Microdomains Stabilize the Postsynapse at the Neuromuscular Junction. EMBO Journal, 25, 4050-4060.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.