EGF Induces Efficient Cx43 Gap Junction Endocytosis in Mouse Embryonic Stem Cell Colonies Via Phosphorylation of Ser262, Ser279/282, and Ser368

Overview

Journal FEBS Lett

Specialty Biochemistry

Date 2014 Feb 5

PMID 24492000

Citations 27

Authors

John T Fong

Wutigri Nimlamool

Matthias M Falk

Affiliations

Soon will be listed here.

Abstract

Gap junctions (GJs) traverse apposing membranes of neighboring cells to mediate intercellular communication by passive diffusion of signaling molecules. We have shown previously that cells endocytose GJs utilizing the clathrin machinery. Endocytosis generates cytoplasmic double-membrane vesicles termed annular gap junctions or connexosomes. However, the signaling pathways and protein modifications that trigger GJ endocytosis are largely unknown. Treating mouse embryonic stem cell colonies - endogenously expressing the GJ protein connexin43 (Cx43) - with epidermal growth factor (EGF) inhibited intercellular communication by 64% and activated both, MAPK and PKC signaling cascades to phosphorylate Cx43 on serines 262, 279/282, and 368. Upon EGF treatment Cx43 phosphorylation transiently increased up to 4-fold and induced efficient (66.4%) GJ endocytosis as evidenced by a 5.9-fold increase in Cx43/clathrin co-precipitation.

Citing Articles

Differential substrate specificity of ERK, JNK, and p38 MAP kinases toward connexin 43.

Latchford L, Perez L, Conage-Pough J, Turk R, Cusimano M, Vargas V J Biol Chem. 2025; 301(3):108178.

PMID: 39798878 PMC: 11870265. DOI: 10.1016/j.jbc.2025.108178.

OXTR-mediated signaling in astrocytes contributes to anxiolysis.

Meinung C, Boi L, Pandamooz S, Mazaud D, Ghezali G, Rouach N Mol Psychiatry. 2024; .

PMID: 39702695 DOI: 10.1038/s41380-024-02870-5.

Acute Adenoviral Infection Elicits an Arrhythmogenic Substrate Prior to Myocarditis.

Padget R, Zeitz M, Blair G, Wu X, North M, Tanenbaum M Circ Res. 2024; 134(7):892-912.

PMID: 38415360 PMC: 11003857. DOI: 10.1161/CIRCRESAHA.122.322437.

Connexin 43 Phosphorylation: Implications in Multiple Diseases.

Zhang M, Wang Z, Chen N Molecules. 2023; 28(13).

PMID: 37446576 PMC: 10343229. DOI: 10.3390/molecules28134914.

Betacellulin regulates gap junction intercellular communication by inducing the phosphorylation of connexin 43 in human granulosa-lutein cells.

Li Y, Chang H, Sung Y, Zhu H, Leung P, Sun Y J Ovarian Res. 2023; 16(1):103.

PMID: 37231448 PMC: 10214552. DOI: 10.1186/s13048-023-01185-3.

References

Fong J, Kells R, Falk M . Two tyrosine-based sorting signals in the Cx43 C-terminus cooperate to mediate gap junction endocytosis. Mol Biol Cell. 2013; 24(18):2834-48. PMC: 3771946. DOI: 10.1091/mbc.E13-02-0111. View

Valiunas V, Polosina Y, Miller H, Potapova I, Valiuniene L, Doronin S . Connexin-specific cell-to-cell transfer of short interfering RNA by gap junctions. J Physiol. 2005; 568(Pt 2):459-68. PMC: 1474730. DOI: 10.1113/jphysiol.2005.090985. View

Gilleron J, Fiorini C, Carette D, Avondet C, Falk M, Segretain D . Molecular reorganization of Cx43, Zo-1 and Src complexes during the endocytosis of gap junction plaques in response to a non-genomic carcinogen. J Cell Sci. 2008; 121(Pt 24):4069-78. DOI: 10.1242/jcs.033373. View

Solan J, Marquez-Rosado L, Sorgen P, Thornton P, Gafken P, Lampe P . Phosphorylation at S365 is a gatekeeper event that changes the structure of Cx43 and prevents down-regulation by PKC. J Cell Biol. 2007; 179(6):1301-9. PMC: 2140020. DOI: 10.1083/jcb.200707060. View

Kanemitsu M, Lau A . Epidermal growth factor stimulates the disruption of gap junctional communication and connexin43 phosphorylation independent of 12-0-tetradecanoylphorbol 13-acetate-sensitive protein kinase C: the possible involvement of mitogen-activated protein.... Mol Biol Cell. 1993; 4(8):837-48. PMC: 300996. DOI: 10.1091/mbc.4.8.837. View

Hesketh G, Shah M, Halperin V, Cooke C, Akar F, Yen T . Ultrastructure and regulation of lateralized connexin43 in the failing heart. Circ Res. 2010; 106(6):1153-63. PMC: 2896878. DOI: 10.1161/CIRCRESAHA.108.182147. View

Park J, Lee M, Heo J, Han H . A potential role of connexin 43 in epidermal growth factor-induced proliferation of mouse embryonic stem cells: involvement of Ca2+/PKC, p44/42 and p38 MAPKs pathways. Cell Prolif. 2008; 41(5):786-802. PMC: 6495952. DOI: 10.1111/j.1365-2184.2008.00552.x. View

Thevenin A, Kowal T, Fong J, Kells R, Fisher C, Falk M . Proteins and mechanisms regulating gap-junction assembly, internalization, and degradation. Physiology (Bethesda). 2013; 28(2):93-116. PMC: 3768091. DOI: 10.1152/physiol.00038.2012. View

Dukes J, Whitley P, Chalmers A . The MDCK variety pack: choosing the right strain. BMC Cell Biol. 2011; 12:43. PMC: 3209442. DOI: 10.1186/1471-2121-12-43. View

10.

Falk M . Connexin-specific distribution within gap junctions revealed in living cells. J Cell Sci. 2000; 113 ( Pt 22):4109-20. DOI: 10.1242/jcs.113.22.4109. View

11.

Berthoud V, Minogue P, Laing J, Beyer E . Pathways for degradation of connexins and gap junctions. Cardiovasc Res. 2004; 62(2):256-67. DOI: 10.1016/j.cardiores.2003.12.021. View

12.

Leach D, Oliphant L . Degradation of annular gap junctions of the equine hoof wall. Acta Anat (Basel). 1984; 120(4):214-9. DOI: 10.1159/000145923. View

13.

Goodenough D, Gilula N . The splitting of hepatocyte gap junctions and zonulae occludentes with hypertonic disaccharides. J Cell Biol. 1974; 61(3):575-90. PMC: 2109309. DOI: 10.1083/jcb.61.3.575. View

14.

Spray D, Burt J . Structure-activity relations of the cardiac gap junction channel. Am J Physiol. 1990; 258(2 Pt 1):C195-205. DOI: 10.1152/ajpcell.1990.258.2.C195. View

15.

Wong R, Pera M, Pebay A . Role of gap junctions in embryonic and somatic stem cells. Stem Cell Rev. 2008; 4(4):283-92. DOI: 10.1007/s12015-008-9038-9. View

16.

Brink P, Valiunas V, Gordon C, Rosen M, Cohen I . Can gap junctions deliver?. Biochim Biophys Acta. 2011; 1818(8):2076-81. DOI: 10.1016/j.bbamem.2011.09.025. View

17.

Boassa D, Solan J, Papas A, Thornton P, Lampe P, Sosinsky G . Trafficking and recycling of the connexin43 gap junction protein during mitosis. Traffic. 2010; 11(11):1471-86. PMC: 3272544. DOI: 10.1111/j.1600-0854.2010.01109.x. View

18.

Falk M, Baker S, Gumpert A, Segretain D, Buckheit 3rd R . Gap junction turnover is achieved by the internalization of small endocytic double-membrane vesicles. Mol Biol Cell. 2009; 20(14):3342-52. PMC: 2710821. DOI: 10.1091/mbc.e09-04-0288. View

19.

Buehr M, Meek S, Blair K, Yang J, Ure J, Silva J . Capture of authentic embryonic stem cells from rat blastocysts. Cell. 2008; 135(7):1287-98. DOI: 10.1016/j.cell.2008.12.007. View

20.

Leithe E, Rivedal E . Epidermal growth factor regulates ubiquitination, internalization and proteasome-dependent degradation of connexin43. J Cell Sci. 2004; 117(Pt 7):1211-20. DOI: 10.1242/jcs.00951. View