title: Nε-lysine acetylation determines dissociation from GAP junctions and lateralization of connexin 43 in normal and dystrophic heart.
abstract: Wanting to explore the epigenetic basis of Duchenne cardiomyopathy, we found that global histone acetylase activity was abnormally elevated and the acetylase P300/CBP-associated factor (PCAF) coimmunoprecipitated with connexin 43 (Cx43), which was N(ε)-lysine acetylated and lateralized in mdx heart. This observation was paralleled by Cx43 dissociation from N-cadherin and zonula occludens 1, whereas pp60-c-Src association was unaltered. In vivo treatment of mdx with the pan-histone acetylase inhibitor anacardic acid significantly reduced Cx43 N(ε)-lysine acetylation and restored its association to GAP junctions (GJs) at intercalated discs. Noteworthy, in normal as well as mdx mice, the class IIa histone deacetylases 4 and 5 constitutively colocalized with Cx43 either at GJs or in the lateralized compartments. The class I histone deacetylase 3 was also part of the complex. Treatment of normal controls with the histone deacetylase pan-inhibitor suberoylanilide hydroxamic acid (MC1568) or the class IIa-selective inhibitor 3-{4-[3-(3-fluorophenyl)-3-oxo-1-propen-1-yl]-1-methyl-1H-pyrrol-2-yl}-N-hydroxy-2-propenamide (MC1568) determined Cx43 hyperacetylation, dissociation from GJs, and distribution along the long axis of ventricular cardiomyocytes. Consistently, the histone acetylase activator pentadecylidenemalonate 1b (SPV106) hyperacetylated cardiac proteins, including Cx43, which assumed a lateralized position that partly reproduced the dystrophic phenotype. In the presence of suberoylanilide hydroxamic acid, cell to cell permeability was significantly diminished, which is in agreement with a Cx43 close conformation in the consequence of hyperacetylation. Additional experiments, performed with Cx43 acetylation mutants, revealed, for the acetylated form of the molecule, a significant reduction in plasma membrane localization and a tendency to nuclear accumulation. These results suggest that Cx43 N(ε)-lysine acetylation may have physiopathological consequences for cell to cell coupling and cardiac function.
authors.affiliation: Laboratorio di Biologia Vascolare e Medicina Rigenerativa, Centro Cardiologico Monzino, 20138 Milan, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Colussi
authors.forename: Jessica
authors.initials: J
authors.lastname: Rosati
authors.forename: Stefania
authors.initials: S
authors.lastname: Straino
authors.forename: Francesco
authors.initials: F
authors.lastname: Spallotta
authors.forename: Roberta
authors.initials: R
authors.lastname: Berni
authors.forename: Donatella
authors.initials: D
authors.lastname: Stilli
authors.forename: Stefano
authors.initials: S
authors.lastname: Rossi
authors.forename: Ezio
authors.initials: E
authors.lastname: Musso
authors.forename: Emilio
authors.initials: E
authors.lastname: Macchi
authors.forename: Antonello
authors.initials: A
authors.lastname: Mai
authors.forename: Gianluca
authors.initials: G
authors.lastname: Sbardella
authors.forename: Sabrina
authors.initials: S
authors.lastname: Castellano
authors.forename: Cristina
authors.initials: C
authors.lastname: Chimenti
authors.forename: Andrea
authors.initials: A
authors.lastname: Frustaci
authors.forename: Angela
authors.initials: A
authors.lastname: Nebbioso
authors.forename: Lucia
authors.initials: L
authors.lastname: Altucci
authors.forename: Maurizio C
authors.initials: MC
authors.lastname: Capogrossi
authors.forename: Carlo
authors.initials: C
authors.lastname: Gaetano
mesh_terms: D000107:Acetylation / Q000187:drug effects; D048729:Anacardic Acids / Q000494:pharmacology; D000818:Animals; D009202:Cardiomyopathies / Q000209:etiology / Q000378:metabolism*; D018031:Connexin 43 / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism*; D051548:Histone Acetyltransferases / Q000378:metabolism; D006877:Hydroxamic Acids; D047468:Immunoprecipitation; D008239:Lysine / Q000378:metabolism*; D051379:Mice; D018101:Mice, Inbred mdx; D008856:Microscopy, Fluorescence; D020388:Muscular Dystrophy, Duchenne / Q000150:complications*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D000077337:Vorinostat; D050880:p300-CBP Transcription Factors / Q000378:metabolism; D000099224:p300-CBP-Associated Factor
chemical_list: D048729:Anacardic Acids; D018031:Connexin 43; D051548:Histone Acetyltransferases; D006877:Hydroxamic Acids; D008239:Lysine; D000077337:Vorinostat; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C543076:GJA1 protein, mouse; C088115:anacardic acid