1. Histone acetyltransferase activity can be modulated by altering the mitochondrial pool of acetyl-CoA.
Histone acetyltransferase activity can be modulated by altering the mitochondrial pool of acetyl-CoA. (A) Total levels of acetyl-CoA were estimated before or after exposure of rho+ or rho0 cells to DM-α-KG using deproteinized samples in a fluorescent-based assay and a standard acetyl-CoA curve. N = 3 biological replicates. (B) HAT activity was estimated in samples from (A) using an in vitro assay. (C) Rho+ and rho0 cells were exposed to the mitochondrial pyruvate kinase inhibitor DCA for 4 h before HAT activity estimation using a fluorescent-based assay; N = 3. (D) Rho+ cells were treated with the mitochondrial citrate carrier inhibitor BTC or an inhibitor of the mitochondrial carnitine transporter, perhexiline (PHX), and HAT assays performed as in (C). (D) Rho+ or rho0 cells were exposed to hydroxycitrate (HC), an inhibitor of the cytosolic ACL, for 4 h. Total acetyl-CoA levels were then estimated using deproteinized samples in a fluorescent-based assay and a standard acetyl-CoA curve. N = 3 biological replicates. (F) HAT activity was gauged in the samples from (E). Statistical analysis for HAT and acetyl-CoA measurements was performed using t test; error bars in all panels represent ±SD.
Oswaldo A Lozoya et al. LSA 2019;2:e
© 2019 Lozoya et al.