CaM kinase II selectively signals to histone deacetylase 4 during cardiomyocyte hypertrophy
Johannes Backs, … , Shurong Chang, Eric N. Olson
Johannes Backs, … , Shurong Chang, Eric N. Olson
Published July 3, 2006
Citation Information: J Clin Invest. 2006;116(7):1853-1864. https://doi.org/10.1172/JCI27438.
View: Text | PDF
Research Article Cardiology

CaM kinase II selectively signals to histone deacetylase 4 during cardiomyocyte hypertrophy

Abstract

Class IIa histone deacetylases (HDACs) regulate a variety of cellular processes, including cardiac growth, bone development, and specification of skeletal muscle fiber type. Multiple serine/threonine kinases control the subcellular localization of these HDACs by phosphorylation of common serine residues, but whether certain class IIa HDACs respond selectively to specific kinases has not been determined. Here we show that calcium/calmodulin-dependent kinase II (CaMKII) signals specifically to HDAC4 by binding to a unique docking site that is absent in other class IIa HDACs. Phosphorylation of HDAC4 by CaMKII promotes nuclear export and prevents nuclear import of HDAC4, with consequent derepression of HDAC target genes. In cardiomyocytes, CaMKII phosphorylation of HDAC4 results in hypertrophic growth, which can be blocked by a signal-resistant HDAC4 mutant. These findings reveal a central role for HDAC4 in CaMKII signaling pathways and have implications for the control of gene expression by calcium signaling in a variety of cell types.

Authors

Johannes Backs, Kunhua Song, Svetlana Bezprozvannaya, Shurong Chang, Eric N. Olson

×

Figure 5

R601 of HDAC4 is required for full responsiveness to CaMKIIδB-T287D.

Options: View larger image (or click on image) Download as PowerPoint
R601 of HDAC4 is required for full responsiveness to CaMKIIδB-T287D. (A ...
(A and B) FLAG-HDAC4 mutants carrying point mutations in the CaMKII docking region were coexpressed with Myc-CaMKIIδB-T287D in COS cells. Coimmunoprecipitation revealed that substitution of R601 with either alanine or phenylalanine prevented a physical interaction with CaMKIIδB-T287D. (C and D) Myc-CaMKIIδB-T287D or CaMKI c.a. and either FLAG-HDAC4-WT, -R601A, or -R601F were coexpressed in COS cells, and HDAC4 localization was determined 1 day after transfection. HDAC4-R601A and -R601F were still responsive to CaMKI but not to CaMKIIδB-T287D and did not colocalize with the kinase. (C) Representative images. Magnification, ×40. (D) Quantitative analysis. (E) Mammalian 2-hybrid assay with GAL4-HDAC4 mutants and VP16–14-3-3. Substitution of R601 with phenylalanine and leucine prevented and with alanine and lysine markedly attenuated 14-3-3 binding. (F) In vitro kinase assay. Active His-CaMKIIδ induced phosphorylation of GST-HDAC4-WT (amino acids 419–670) but not of the GST-HDAC4-R601F (419–670) mutant. Ca2+-depletion by EGTA prevented HDAC4 phosphorylation by CaMKII. (G) GST pull-down assay with GST, GST-HDAC4-WT (419–670), GST-HDAC4-R601F (419–670) mutant, and active His-CaMKIIδ. (H) Coimmunoprecipitation revealed that HA-CaMKI docks to FLAG-HDAC4-WT and -R601F to the same degree, however, it docks to HDAC5 with greater affinity.