Neuronal cell depolarization induces intragenic chromatin modifications affecting NCAM alternative splicing.
In search for physiological pathways affecting alternative splicing through its kinetic coupling with transcription, we found that membrane depolarization of neuronal cells triggers the skipping of exon 18 from the neural cell adhesion molecule (NCAM) mRNA, independently of the calcium/calmodulin protein kinase IV pathway. We show that this exon responds to RNA polymerase II elongation, because its inclusion is increased by a slow polymerase II mutant. Depolarization affects the chromatin template in a specific way, by causing H3K9 hyper-acetylation restricted to an internal region of the NCAM gene surrounding the alternative exon. This intragenic histone hyper-acetylation is not paralleled by acetylation at the promoter, is associated with chromatin relaxation, and is linked to H3K36 tri-methylation. The effects on acetylation and splicing fully revert when the depolarizing conditions are withdrawn and can be both duplicated and potentiated by the histone deacetylase inhibitor trichostatin A. Our results are consistent with a mechanism involving the kinetic coupling of splicing and transcription in response to depolarization through intragenic epigenetic changes on a gene that is relevant for the differentiation and function of neuronal cells.
Schor
Ignacio E
IE
Departamento de Fisiología, Biología Molecular y Celular, IFIBYNE-UBA Consejo Nacional de Investigaciones Científicas y Técnicas of Argentina, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.
Rascovan
Nicolás
N
Pelisch
Federico
F
Alló
Mariano
M
Kornblihtt
Alberto R
AR
D000107:Acetylation; D017398:Alternative Splicing; D000818:Animals; D002843:Chromatin / Q000235:genetics*; D044127:Epigenesis, Genetic; D005091:Exons; D006657:Histones / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology*; D019006:Neural Cell Adhesion Molecules / Q000235:genetics*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D012319:RNA Polymerase II / Q000378:metabolism; D051381:Rats
D002843:Chromatin; D006657:Histones; D019006:Neural Cell Adhesion Molecules; D012319:RNA Polymerase II