Cx43 phosphorylation-mediated effects on ERK and Akt protect against ischemia reperfusion injury and alter the stability of the stress-inducible protein NDRG1.

Abstract

Gap junctions contain intercellular channels that enable intercellular communication of small molecules while also serving as a signaling scaffold. Connexins, the proteins that form gap junctions in vertebrates, are highly regulated and typically have short (<2 h) half-lives. Connexin43 (Cx43), the predominate connexin in the myocardium and epithelial tissues, is phosphorylated on more than a dozen serine residues and interacts with a variety of protein kinases. These interactions regulate Cx43 and gap junction formation and stability. Casein kinase 1 (CK1)-mediated phosphorylation of Cx43 promotes gap junction assembly. Using murine knock-in technology and quantitative PCR, immunoblotting, and immunoprecipitation assays, we show here that mutation of the CK1 phosphorylation sites in Cx43 reduces the levels of total Cx43 in the myocardium and increases Cx43 phosphorylation on sites phosphorylated by extracellular signal-regulated kinase (ERK). In aged myocardium, we found that, compared with WT Cx43, mutant Cx43 expression increases ERK activation, phosphorylation of Akt substrates, and protection from ischemia-induced injury. Our findings also uncovered that Cx43 interacts with the hypoxia-inducible protein N-Myc downstream-regulated gene 1 protein (NDRG1) and that Cx43 phosphorylation status controls this interaction and dramatically affects NDRG1 stability. We propose that, in addition to altering gap junction stability, Cx43 phosphorylation directly and dynamically regulates cellular signaling through ERK and Akt in response to ischemic injury. We conclude that gap junction-dependent NDRG1 regulation might explain some cellular responses to hypoxia.

• Authors: Joell L Solan, Lucrecia Márquez-Rosado, Paul D Lampe
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 294 Issue 31 Pg. 11762-11771 (08 02 2019) ISSN: 1083-351X [Electronic] United States
• PMID: 31189653 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2019 Solan et al.

Chemical References

• Cell Cycle Proteins
• Connexin 43
• Intracellular Signaling Peptides and Proteins
• N-myc downstream-regulated gene 1 protein
• Casein Kinase I
• Proto-Oncogene Proteins c-akt
• Extracellular Signal-Regulated MAP Kinases

Topics

• Aging
• Animals
• Casein Kinase I (metabolism)
• Cell Cycle Proteins (metabolism)
• Connexin 43 (genetics, metabolism)
• Dogs
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Gap Junctions (metabolism)
• Intracellular Signaling Peptides and Proteins (metabolism)
• Madin Darby Canine Kidney Cells
• Mice
• Mice, Inbred C57BL
• Mutagenesis
• Myocardium (metabolism)
• Phosphorylation (drug effects)
• Protein Stability
• Proto-Oncogene Proteins c-akt (metabolism)
• Reperfusion Injury (metabolism, pathology)
• Substrate Specificity