Gap junction turnover, intracellular trafficking, and phosphorylation of connexin43 in brefeldin A-treated rat mammary tumor cells

doi:10.1083/jcb.131.5.1193

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Gap junction turnover, intracellular trafficking, and phosphorylation of connexin43 in brefeldin A-treated rat mammary tumor cells

DW Laird, M Castillo and L Kasprzak
Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada.

Intercellular gap junction channels are thought to form when oligomers of connexins from one cell (connexons) register and pair with connexons from a neighboring cell en route to forming tightly packed arrays (plaques). In the current study we used the rat mammary BICR-M1Rk tumor cell line to examine the trafficking, maturation, and kinetics of connexin43 (Cx43). Cx43 was conclusively shown to reside in the Golgi apparatus in addition to sites of cell-cell apposition in these cells and in normal rat kidney cells. Brefeldin A (BFA) blocked Cx43 trafficking to the surface of the mammary cells and also prevented phosphorylation of the 42-kD form of Cx43 to 44- and 46-kD species. However, phosphorylation of Cx43 occurred in the presence of BFA while it was still a resident of the ER or Golgi apparatus yielding a 43-kD form of Cx43. Moreover, the 42- and 43-kD forms of Cx43 trapped in the ER/Golgi compartment were available for gap junction assembly upon the removal of BFA. Mammary cells treated with BFA for 6 h lost preexisting gap junction "plaques," as well as the 44- and 46-kD forms of Cx43 and functional coupling. These events were reversible 1 h after the removal of BFA and not dependent on protein synthesis. In summary, we provide strong evidence that in BICR-M1Rk tumor cells: (a) Cx43 is transiently phosphorylated in the ER/Golgi apparatus, (b) Cx43 trapped in the ER/Golgi compartment is not subject to rapid degradation and is available for the assembly of new gap junction channels upon the removal of BFA, (c) the rapid turnover of gap junction plaques is correlated with the loss of the 44- and 46-kD forms of Cx43.
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Flenniken, A. M., Osborne, L. R., Anderson, N., Ciliberti, N., Fleming, C., Gittens, J. E. I., Gong, X.-Q., Kelsey, L. B., Lounsbury, C., Moreno, L., Nieman, B. J., Peterson, K., Qu, D., Roscoe, W., Shao, Q., Tong, D., Veitch, G. I. L., Voronina, I., Vukobradovic, I., Wood, G. A., Zhu, Y., Zirngibl, R. A., Aubin, J. E., Bai, D., Bruneau, B. G., Grynpas, M., Henderson, J. E., Henkelman, R. M., McKerlie, C., Sled, J. G., Stanford, W. L., Laird, D. W., Kidder, G. M., Adamson, S. L., Rossant, J. (2005). A Gja1 missense mutation in a mouse model of oculodentodigital dysplasia. Development 132: 4375-4386 [Abstract] [Full Text]
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Lauf, U., Giepmans, B. N. G., Lopez, P., Braconnot, S., Chen, S.-C., Falk, M. M. (2002). Dynamic trafficking and delivery of connexons to the plasma membrane and accretion to gap junctions in living cells. Proc. Natl. Acad. Sci. USA 99: 10446-10451 [Abstract] [Full Text]
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VanSlyke, J. K., Musil, L. S. (2002). Dislocation and degradation from the ER are regulated by cytosolic stress. JCB 157: 381-394 [Abstract] [Full Text]
Anderson, S. C., Stone, C., Tkach, L., SundarRaj, N. (2002). Rho and Rho-Kinase (ROCK) Signaling in Adherens and Gap Junction Assembly in Corneal Epithelium. IOVS 43: 978-986 [Abstract] [Full Text]
Das Sarma, J., Meyer, R. A., Wang, F., Abraham, V., Lo, C. W., Koval, M. (2002). Multimeric connexin interactions prior to the trans-Golgi network. J. Cell Sci. 114: 4013-4024 [Abstract] [Full Text]
Lampe, P. D., Qiu, Q., Meyer, R. A., TenBroek, E. M., Walseth, T. F., Starich, T. A., Grunenwald, H. L., Johnson, R. G. (2001). Gap junction assembly: PTX-sensitive G proteins regulate the distribution of connexin43 within cells. Am. J. Physiol. Cell Physiol. 281: C1211-C1222 [Abstract] [Full Text]
Lin, R., Warn-Cramer, B. J., Kurata, W. E., Lau, A. F. (2001). v-Src phosphorylation of connexin 43 on Tyr247 and Tyr265 disrupts gap junctional communication. JCB 154: 815-828 [Abstract] [Full Text]
Le, A.-C. N., Musil, L. S. (2001). A novel role for FGF and extracellular signal-regulated kinase in gap junction-mediated intercellular communication in the lens. JCB 154: 197-216 [Abstract] [Full Text]
Woodward, T. L., Mienaltowski, A. S., Modi, R. R., Bennett, J. M., Haslam, S. Z. (2001). Fibronectin and the {{alpha}}5{beta}1 Integrin Are Under Developmental and Ovarian Steroid Regulation in the Normal Mouse Mammary Gland. Endocrinology 142: 3214-3222 [Abstract] [Full Text]
Martin, P. E. M., Blundell, G., Ahmad, S., Errington, R. J., Evans, W. H. (2001). Multiple pathways in the trafficking and assembly of connexin 26, 32 and 43 into gap junction intercellular communication channels. J. Cell Sci. 114: 3845-3855 [Abstract] [Full Text]
Jordan, K, Chodock, R, Hand, A., Laird, D. (2001). The origin of annular junctions: a mechanism of gap junction internalization. J. Cell Sci. 114: 763-773 [Abstract]
Quist, A. P., Rhee, S. K., Lin, H., Lal, R. (2000). Physiological Role of Gap-Junctional Hemichannels: Extracellular Calcium-Dependent Isosmotic Volume Regulation. JCB 148: 1063-1074 [Abstract] [Full Text]
Doble, B. W., Ping, P., Kardami, E. (2000). The {epsilon} Subtype of Protein Kinase C Is Required for Cardiomyocyte Connexin-43 Phosphorylation. Circ. Res. 86: 293-301 [Abstract] [Full Text]
Falk, M. (2000). Connexin-specific distribution within gap junctions revealed in living cells. J. Cell Sci. 113: 4109-4120 [Abstract]
Paulson, A., Lampe, P., Meyer, R., TenBroek, E, Atkinson, M., Walseth, T., Johnson, R. (2000). Cyclic AMP and LDL trigger a rapid enhancement in gap junction assembly through a stimulation of connexin trafficking. J. Cell Sci. 113: 3037-3049 [Abstract]
Athanassiou, M., Hu, Y., Jing, L., Houle, B., Zarbl, H., Mikheev, A. M. (1999). Stabilization and Reactivation of the p53 Tumor Suppressor Protein in Nontumorigenic Revertants of HeLa Cervical Cancer Cells. Cell Growth Differ. 10: 729-737 [Abstract] [Full Text]
Cyr, D. G., Hermo, L., Egenberger, N., Mertineit, C., Trasler, J. M., Laird, D. W. (1999). Cellular Immunolocalization of Occludin during Embryonic and Postnatal Development of the Mouse Testis and Epididymis. Endocrinology 140: 3815-3825 [Abstract] [Full Text]
Laird, D. W., Fistouris, P., Batist, G., Alpert, L., Huynh, H. T., Carystinos, G. D., Alaoui-Jamali, M. A. (1999). Deficiency of Connexin43 Gap Junctions Is an Independent Marker for Breast Tumors. Cancer Res. 59: 4104-4110 [Abstract] [Full Text]
Guo, Y., Martinez-Williams, C., Gilbert, K. A., Rannels, D. E. (1999). Inhibition of gap junction communication in alveolar epithelial cells by 18alpha -glycyrrhetinic acid. Am. J. Physiol. Lung Cell. Mol. Physiol. 276: L1018-L1026 [Abstract] [Full Text]
Jordan, K., Solan, J. L., Dominguez, M., Sia, M., Hand, A., Lampe, P. D., Laird, D. W. (1999). Trafficking, Assembly, and Function of a Connexin43-Green Fluorescent Protein Chimera in Live Mammalian Cells. Mol. Biol. Cell 10: 2033-2050 [Abstract] [Full Text]
George, C. H., Kendall, J. M., Evans, W. H. (1999). Intracellular Trafficking Pathways in the Assembly of Connexins into Gap Junctions. J. Biol. Chem. 274: 8678-8685 [Abstract] [Full Text]
Carystinos, G. D., Katabi, M. M., Laird, D. W., Galipeau, J., Chan, H., Alaoui-Jamali, M. A., Batist, G. (1999). Cyclic-AMP Induction of Gap Junctional Intercellular Communication Increases Bystander Effect in Suicide Gene Therapy. Clin. Cancer Res. 5: 61-68 [Abstract] [Full Text]
Lampe, P. D., Nguyen, B. P., Gil, S., Usui, M., Olerud, J., Takada, Y., Carter, W. G. (1998). Cellular Interaction of Integrin {alpha}3{beta}1 with Laminin 5 Promotes Gap Junctional Communication. JCB 143: 1735-1747 [Abstract] [Full Text]
George, C. H., Kendall, J. M., Campbell, A. K., Evans, W. H. (1998). Connexin-Aequorin Chimerae Report Cytoplasmic Calcium Environments along Trafficking Pathways Leading to Gap Junction Biogenesis in Living COS-7 Cells. J. Biol. Chem. 273: 29822-29829 [Abstract] [Full Text]
Nadarajah, B., Makarenkova, H., Becker, D. L., Evans, W. H., Parnavelas, J. G. (1998). Basic FGF Increases Communication between Cells of the Developing Neocortex. J. Neurosci. 18: 7881-7890 [Abstract] [Full Text]
Peters, N. S., Wit, A. L. (1998). Myocardial Architecture and Ventricular Arrhythmogenesis. Circulation 97: 1746-1754 [Full Text]
Falk, M. M., Gilula, N. B. (1998). Connexin Membrane Protein Biosynthesis Is Influenced by Polypeptide Positioning within the Translocon and Signal Peptidase Access. J. Biol. Chem. 273: 7856-7864 [Abstract] [Full Text]
Woodward, T., Sia, M., Blaschuk, O., Turner, J., Laird, D. (1998). Deficient epithelial-fibroblast heterocellular gap junction communication can be overcome by co-culture with an intermediate cell type but not by E-cadherin transgene expression. J. Cell Sci. 111: 3529-3539 [Abstract]
Viger, R., Mertineit, C, Trasler, J., Nemer, M (1998). Transcription factor GATA-4 is expressed in a sexually dimorphic pattern during mouse gonadal development and is a potent activator of the Mullerian inhibiting substance promoter. Development 125: 2665-2675 [Abstract]
Mertineit, C, Yoder, J., Taketo, T, Laird, D., Trasler, J., Bestor, T. (1998). Sex-specific exons control DNA methyltransferase in mammalian germ cells. Development 125: 889-897 [Abstract]
Sakakibara, A., Furuse, M., Saitou, M., Ando-Akatsuka, Y., Tsukita, S. (1997). Possible Involvement of Phosphorylation of Occludin in Tight Junction Formation. JCB 137: 1393-1401 [Abstract] [Full Text]
Xie, H.-q., Laird, D. W., Chang, T.-H., Hu, V. W. (1997). A Mitosis-specific Phosphorylation of the Gap Junction Protein Connexin43 in Human Vascular Cells: Biochemical Characterization and Localization. JCB 137: 203-210 [Abstract] [Full Text]
Musil, L. S., Le, A.-C. N., VanSlyke, J. K., Roberts, L. M. (2000). Regulation of Connexin Degradation as a Mechanism to Increase Gap Junction Assembly and Function. J. Biol. Chem. 275: 25207-25215 [Abstract] [Full Text]
VanSlyke, J. K., Musil, L. S. (2002). Dislocation and degradation from the ER are regulated by cytosolic stress. JCB 157: 381-394 [Abstract] [Full Text]