Beta-Adrenergic Activation Enhances Histone Deacetylase 4 nuclear Localization via Pka and Counters Camkii-Dependent Effects in Adult Rabbit Cardiac Myocytes

Helmstadter, Kathryn; Erickson, Jeffrey R.; Dao, Khanha; Bossuyt, Julie; Bers, Donald M.

doi:10.1016/j.bpj.2010.12.646

Cited by 2 publications

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“…Phosphorylation of HDAC5 at serine 280 by PKA interrupts the association of HDAC5 with 14–3–3 and thereby inhibits nuclear export of HDAC5 and promotes nuclear retention (Ha et al 2010; Chang et al 2013). An alignment analysis shows that there is a potential PKA phosphorylation site at serine 265 and/or 266 in HDAC4 (D. Bers, personal communication; Helmstadter et al 2011), equivalent to serine 280 in HDAC5. Therefore, to pursue the mechanism of nuclear accumulation of HDAC4 by cAMP we tested the effects of beta‐adrenergic activation on HDAC4 (S265/266A)‐GFP (Helmstadter et al 2011), an HDAC4‐GFP mutant construct having serines 265 and 266 replaced with alanines, which eliminates the possibility of phosphorylation of these residues.…”

Section: Resultsmentioning

confidence: 99%

“…An alignment analysis shows that there is a potential PKA phosphorylation site at serine 265 and/or 266 in HDAC4 (D. Bers, personal communication; Helmstadter et al 2011), equivalent to serine 280 in HDAC5. Therefore, to pursue the mechanism of nuclear accumulation of HDAC4 by cAMP we tested the effects of beta‐adrenergic activation on HDAC4 (S265/266A)‐GFP (Helmstadter et al 2011), an HDAC4‐GFP mutant construct having serines 265 and 266 replaced with alanines, which eliminates the possibility of phosphorylation of these residues. Under control resting conditions, the nuclear/cytoplasmic fluorescence ratio in fibres expressing HDAC4 (S265/266A)‐GFP was not significantly different from fibres expressing wild‐type (wt) HDAC4‐GFP (1.86 ± 0.29 in 14 nuclei from 8 fibres expressing HDAC4‐GFP vs .…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant adenovirus (Ad5) containing HDAC4-GFP was produced as described previously (Liu et al 2005) according to the methods of Hardy et al (1997) with a cytomegalovirus (CMV) promoter. Recombinant adenovirus expressing HDAC4-GFP S265/266A was constructed with an AdEasy Adenoviral Vector System according to the manufacturer's instruction (Agilent Technologies, Santa Clara, CA, USA) with a CMV promoter and was a gift from Dr D. M. Bers (University of California Davis; Helmstadter et al 2011). Virus infections were performed with about 10 8 particles per muscle fibre.…”

Section: Infection Of Recombinant Adenoviruses In Muscle Fibresmentioning

confidence: 99%

“…In contrast, phosphorylation by protein kinase A (PKA) causes nuclear accumulation of HDACs in C2C12 myoblasts and vascular smooth muscle cells (Du et al 2008; Gordon et al 2009). Recent studies show that PKA phosphorylates HDAC5 at serine 280 (Ha et al 2010; Chang et al 2013), or HDAC4 at serine 265/266 (Helmstadter et al 2011), which is localized between the phosphorylatable 14–3–3 binding sites critical for nuclear export of HDAC4 or 5. Phosphorylation of serine 280 of HDAC5 by PKA interrupts the binding of 14–3–3 without hindering phosphorylation at serine 259 and 498 by other kineses (Ha et al 2010; Chang et al 2013).…”

Section: Introductionmentioning

confidence: 99%

“…Phosphorylation of serine 280 of HDAC5 by PKA interrupts the binding of 14–3–3 without hindering phosphorylation at serine 259 and 498 by other kineses (Ha et al 2010; Chang et al 2013). Thus, it is anticipated that phosphorylation by PKA or CaMKII should produce antagonistic effects on nuclear/cytoplasmic distribution of class IIa HDACs, and such antagonistic effects have recently been observed in adult cardiomyocytes (Helmstadter et al 2011; D. Bers, personal communication).…”

Section: Introductionmentioning

confidence: 99%

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Opposing HDAC4 nuclear fluxes due to phosphorylation by β‐adrenergic activated protein kinase A or by activity or Epac activated CaMKII in skeletal muscle fibres

Schneider

2013

The Journal of Physiology

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Key points• Application of either the beta-adrenergic agonist isoproterenol, dibutyryl cAMP or specific PKA activator N6 benzoyl cAMP caused nuclear influx of wild-type (wt) HDAC4-GFP expressed in cultured adult skeletal muscle fibres, but caused no change in nuclear/cytoplasmic distribution of expressed 'mut' HDAC4-GFP mutated (S 265 and 266 to A) at the protein kinase A (PKA) phosphorylation site(s), demonstrating that PKA promotes HDAC4 nuclear influx by phosphorylation of HDAC4 at the PKA sites.• In non-transfected muscle fibres, myocyte enhancer factor 2 (MEF2)-driven luc reporter activity was decreased by application of isoproterenol, indicating that endogenous HDAC4 increased in fibre nuclei and suppressed MEF2 transcriptional activity. Levels of phosphorylated (i.e. active) PKA were elevated by exposure to dibutyryl (Db) cAMP.• Fibre repetitive electrical stimulation with 10 Hz trains caused a CaMKII dependent nuclear efflux of wt and mut HDAC4, which was partially decreased by Db cAMP for wt but not for mut HDAC4-GFP.• The specific activator 8-CPT of Epac caused efflux of both wt and mut HDAC4-GFP, which was eliminated by the CaMK inhibitor KN-93 or by buffering cytosolic Ca 2+ using BAPTA-AM loading, both of which also eliminated a slow elevation of cytosolic Ca 2+ during 8-CPT application.• Using a submaximally effective stimulus frequency of 4 Hz trains of electrical stimulation, Db cAMP increased the rate of nuclear influx of mut HDAC4-GFP, which cannot be phosphorylated by PKA but can be phosphorylated by CaMKII, which is here activated via the cAMP/Epac pathway.• Immunostain for active PKA or for GTP-bound RAP1, which is an indicator of Epac activation, showed responses consistent with the functional results above.Abstract Class IIa histone deacetylases (HDACs) move between skeletal muscle fibre cytoplasm and nuclei in response to various stimuli, suppressing activity of the exclusively nuclear transcription factor Mef2. Protein kinase A (PKA) phosphorylates class IIa HDACs in cardiac muscle, resulting in HDAC nuclear accumulation, but this has not been examined in skeletal muscle. Using HDAC4-green fluorescent protein (HDAC4-GFP) expressed in isolated skeletal muscle fibres, we now show that activation of PKA by the beta-receptor agonist isoproterenol or dibutyryl (Db) cAMP causes a steady HDAC4-GFP nuclear influx. The beta-receptor blocker propranolol or PKA inhibitor Rp-cAMPS blocks the effects of isoproterenol on the nuclear influx of HDAC4-GFP, and Rp-cAMPS blocks the effects of Db cAMP. The HDAC4-GFP construct having serines 265 and 266 replaced with alanines, HDAC4 (S265/266A)-GFP, did not respond

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Infection Of Recombinant Adenoviruses In Muscle Fibresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Opposing HDAC4 nuclear fluxes due to phosphorylation by β‐adrenergic activated protein kinase A or by activity or Epac activated CaMKII in skeletal muscle fibres

Schneider

2013

The Journal of Physiology

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CaMKII and PKA-dependent phosphorylation co-regulate nuclear localization of HDAC4 in adult cardiomyocytes

et al. 2021

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Nuclear histone deacetylase 4 (HDAC4) represses MEF2-mediated transcription, implicated in the development of heart failure. CaMKII-dependent phosphorylation drives nucleus-to-cytoplasm HDAC4 shuttling, but protein kinase A (PKA) is also linked to HDAC4 translocation. However, the interplay of CaMKII and PKA in regulating adult cardiomyocyte HDAC4 translocation is unclear. Here we sought to determine the interplay of PKA- and CaMKII-dependent HDAC4 phosphorylation and translocation in adult mouse, rabbit and human ventricular myocytes. Confocal imaging and protein analyses revealed that inhibition of CaMKII—but not PKA, PKC or PKD—raised nucleo-to-cytoplasmic HDAC4 fluorescence ratio (FNuc/FCyto) by ~ 50%, indicating baseline CaMKII activity that limits HDAC4 nuclear localization. Further CaMKII activation (via increased extracellular [Ca2+], high pacing frequencies, angiotensin II or overexpression of CaM or CaMKIIδC) led to significant HDAC4 nuclear export. In contrast, PKA activation by isoproterenol or forskolin drove HDAC4 into the nucleus (raising FNuc/FCyto by > 60%). These PKA-mediated effects were abolished in cells pretreated with PKA inhibitors and in cells expressing mutant HDAC4 in S265/266A mutant. In physiological conditions where both kinases are active, PKA-dependent nuclear accumulation of HDAC4 was predominant in the very early response, while CaMKII-dependent HDAC4 export prevailed upon prolonged stimuli. This orchestrated co-regulation was shifted in failing cardiomyocytes, where CaMKII-dependent effects predominated over PKA-dependent response. Importantly, human cardiomyocytes showed similar CaMKII- and PKA-dependent HDAC4 shifts. Collectively, CaMKII limits nuclear localization of HDAC4, while PKA favors HDAC4 nuclear retention and S265/266 is essential for PKA-mediated regulation. These pathways thus compete in HDAC4 nuclear localization and transcriptional regulation in cardiac signaling.

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Beta-Adrenergic Activation Enhances Histone Deacetylase 4 nuclear Localization via Pka and Counters Camkii-Dependent Effects in Adult Rabbit Cardiac Myocytes

Cited by 2 publications

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Opposing HDAC4 nuclear fluxes due to phosphorylation by β‐adrenergic activated protein kinase A or by activity or Epac activated CaMKII in skeletal muscle fibres

Opposing HDAC4 nuclear fluxes due to phosphorylation by β‐adrenergic activated protein kinase A or by activity or Epac activated CaMKII in skeletal muscle fibres

CaMKII and PKA-dependent phosphorylation co-regulate nuclear localization of HDAC4 in adult cardiomyocytes

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