Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes
| Autor: | Friederike Cuello, Alexandra J. Candasamy, Elisabeth Ehler, Metin Avkiran, Elizabeth M. Kemp, Konstantina Stathopoulou, Robert S. Haworth |
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| Rok vydání: | 2014 |
| Předmět: |
CaMK
Ca2+/calmodulin-dependent protein kinase Muscle Proteins HDAC histone deacetylase Biochemistry Mice 0302 clinical medicine cMyBP-C cardiac myosin-binding protein C Myocytes Cardiac Phosphorylation Cells Cultured Protein Kinase C MOI multiplicity of infection 0303 health sciences Histone deacetylase 5 Endothelin-1 MEF2 Transcription Factors Kinase Intracellular Signaling Peptides and Proteins protein kinase LIM Domain Proteins musculoskeletal system 3. Good health Isoenzymes PKD protein kinase D FHL four-and-a-half LIM domains cardiovascular system caPKD constitutively active catalytic domain of PKD TAC transverse aortic constriction Signal transduction signal transduction Research Article Mef2 NRVM neonatal rat ventricular myocyte Heart Ventricles Recombinant Fusion Proteins LIM-Homeodomain Proteins cardiac myocyte Biology ERK extracellular-signal-regulated kinase Histone Deacetylases neurohormonal stimulation MEF2 myocyte enhancer factor 2 03 medical and health sciences IVK in vitro kinase cTnI inhibitory subunit of cardiac troponin PKC protein kinase C Animals Humans Protein kinase A Molecular Biology CRM1 chromosome region maintenance 1 four-and-a-half LIM (FHL) ARVM adult rat ventricular myocyte 030304 developmental biology PE phenylephrine pfu plaque-forming unit ET1 endothelin 1 Cell Biology BPKDi bipyridyl PKD inhibitor Molecular biology Peptide Fragments FHL1 Rats FHL2 Enzyme Activation Animals Newborn histone deacetylase MuRF muscle RING finger Protein Processing Post-Translational 030217 neurology & neurosurgery Transcription Factors |
| Zdroj: | Biochemical Journal |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj20131026 |
| Popis: | PKD (protein kinase D) is a serine/threonine kinase implicated in multiple cardiac roles, including the phosphorylation of the class II HDAC5 (histone deacetylase isoform 5) and thereby de-repression of MEF2 (myocyte enhancer factor 2) transcription factor activity. In the present study we identify FHL1 (four-and-a-half LIM domains protein 1) and FHL2 as novel binding partners for PKD in cardiac myocytes. This was confirmed by pull-down assays using recombinant GST-fused proteins and heterologously or endogenously expressed PKD in adult rat ventricular myocytes or NRVMs (neonatal rat ventricular myocytes) respectively, and by co-immunoprecipitation of FHL1 and FHL2 with GFP–PKD1 fusion protein expressed in NRVMs. In vitro kinase assays showed that neither FHL1 nor FHL2 is a PKD1 substrate. Selective knockdown of FHL1 expression in NRVMs significantly inhibited PKD activation and HDAC5 phosphorylation in response to endothelin 1, but not to the α1-adrenoceptor agonist phenylephrine. In contrast, selective knockdown of FHL2 expression caused a significant reduction in PKD activation and HDAC5 phosphorylation in response to both stimuli. Interestingly, neither intervention affected MEF2 activation by endothelin 1 or phenylephrine. We conclude that FHL1 and FHL2 are novel cardiac PKD partners, which differentially facilitate PKD activation and HDAC5 phosphorylation by distinct neurohormonal stimuli, but are unlikely to regulate MEF2-driven transcriptional reprogramming. Protein kinase D has multiple roles in cardiac myocytes, where its regulatory mechanisms remain incompletely defined. In the present study we identify four-and-a-half LIM domains proteins 1 and 2 as novel binding partners and regulators of protein kinase D in this cell type. |
| Databáze: | OpenAIRE |
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