Class IIa Histone Deacetylases Are Conserved Regulators of Circadian Function*

Background: Class IIa HDACs are signal-dependent transcriptional corepressors that regulate cell differentiation programs and liver gluconeogenesis.Results: HDAC5 influences BMAL1 acetylation and interfering with normal expression levels of class IIa HDACs disrupts circadian rhythms.Conclusion: Class IIa HDACs regulate the robustness of cellular clocks and behavioral activity rhythms.Significance: Class IIa HDACs provide a conserved link between circadian clocks and metabolic signaling pathways.

Protocol publication

[…] To assess the effect of HDAC inhibitors on BMAL1 acetylation, NIH3T3 cells transfected with BMAL1-Myc using FuGENE 6 (Roche Applied Science) were treated with indicated HDAC inhibitors 24 h after transfection. After 18 h, cells were lysed in hot SDS sample buffer. To assess the effect of HDAC5 expression on BMAL1 acetylation protein extracts were prepared from cells 40 h after transfection. Proteins were resolved by SDS-PAGE using pre-cast Bis-Tris gels (Invitrogen) and immunoblotted using standard protocols. Band intensities were quantified with ImageJ software. Statistical significance was determined with a two-tailed paired Student's t test using the SPSS Statistical suite (IBM). […]