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Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes

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Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes. / Stathopoulou, Konstantina; Cuello, Friederike; Candasamy, Alexandra J; Kemp, Elizabeth M; Ehler, Elisabeth; Haworth, Robert S; Avkiran, Metin.

In: BIOCHEM J, Vol. 457, No. 3, 01.02.2014, p. 451-461.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Stathopoulou, K, Cuello, F, Candasamy, AJ, Kemp, EM, Ehler, E, Haworth, RS & Avkiran, M 2014, 'Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes', BIOCHEM J, vol. 457, no. 3, pp. 451-461. https://doi.org/10.1042/BJ20131026

APA

Stathopoulou, K., Cuello, F., Candasamy, A. J., Kemp, E. M., Ehler, E., Haworth, R. S., & Avkiran, M. (2014). Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes. BIOCHEM J, 457(3), 451-461. https://doi.org/10.1042/BJ20131026

Vancouver

Stathopoulou K, Cuello F, Candasamy AJ, Kemp EM, Ehler E, Haworth RS et al. Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes. BIOCHEM J. 2014 Feb 1;457(3):451-461. https://doi.org/10.1042/BJ20131026

Bibtex

@article{fa7a134352314d7b8858ad5f36eb71c5,
title = "Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes",
abstract = "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.",
keywords = "Animals, Animals, Newborn, Cells, Cultured, Endothelin-1, Enzyme Activation, Heart Ventricles, Histone Deacetylases, Humans, Intracellular Signaling Peptides and Proteins, Isoenzymes, LIM Domain Proteins, LIM-Homeodomain Proteins, MEF2 Transcription Factors, Mice, Muscle Proteins, Myocytes, Cardiac, Peptide Fragments, Phosphorylation, Protein Kinase C, Protein Processing, Post-Translational, Rats, Recombinant Fusion Proteins, Transcription Factors",
author = "Konstantina Stathopoulou and Friederike Cuello and Candasamy, {Alexandra J} and Kemp, {Elizabeth M} and Elisabeth Ehler and Haworth, {Robert S} and Metin Avkiran",
year = "2014",
month = feb,
day = "1",
doi = "10.1042/BJ20131026",
language = "English",
volume = "457",
pages = "451--461",
journal = "BIOCHEM J",
issn = "0264-6021",
publisher = "PORTLAND PRESS LTD",
number = "3",

}

RIS

TY - JOUR

T1 - Four-and-a-half LIM domains proteins are novel regulators of the protein kinase D pathway in cardiac myocytes

AU - Stathopoulou, Konstantina

AU - Cuello, Friederike

AU - Candasamy, Alexandra J

AU - Kemp, Elizabeth M

AU - Ehler, Elisabeth

AU - Haworth, Robert S

AU - Avkiran, Metin

PY - 2014/2/1

Y1 - 2014/2/1

N2 - 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.

AB - 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.

KW - Animals

KW - Animals, Newborn

KW - Cells, Cultured

KW - Endothelin-1

KW - Enzyme Activation

KW - Heart Ventricles

KW - Histone Deacetylases

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - Isoenzymes

KW - LIM Domain Proteins

KW - LIM-Homeodomain Proteins

KW - MEF2 Transcription Factors

KW - Mice

KW - Muscle Proteins

KW - Myocytes, Cardiac

KW - Peptide Fragments

KW - Phosphorylation

KW - Protein Kinase C

KW - Protein Processing, Post-Translational

KW - Rats

KW - Recombinant Fusion Proteins

KW - Transcription Factors

U2 - 10.1042/BJ20131026

DO - 10.1042/BJ20131026

M3 - SCORING: Journal article

C2 - 24219103

VL - 457

SP - 451

EP - 461

JO - BIOCHEM J

JF - BIOCHEM J

SN - 0264-6021

IS - 3

ER -