The role of p38 signaling and poly(ADP-ribosyl)ation-induced metabolic collapse in the osteogenic differentiation-coupled cell death pathway

Agnieszka Robaszkiewicz, Zsuzsanna Valkó, Katalin Kovács, Csaba Hegedus, Edina Bakondi, P. Bai, L. Virag

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Osteogenic differentiation is a multistep process regulated by a diverse set of morphogenic and transcription factors. Previously we identified endogenous hydrogen peroxide-induced poly(ADP-ribose) polymerase-1 (PARP1) activation as a mediator of osteodifferentiation and associated cell death. Here we set out to investigate whether or not activation of PARP1 is dependent on DNA breaks and how PARP1 mediates cell death during osteodifferentiation of mesenchymal stem cells and SAOS-2 cells. Here we show that the MAP kinases p38, JNK, and ERK1/2 become activated during the differentiation process. However, only p38 activation depended both on hydrogen peroxide production and on PARP1 activation as the hydrogen peroxide decomposing enzyme catalase, the PARP inhibitor PJ34, and the silencing of PARP1 suppressed p38 activation. Inhibition of p38 suppressed cell death and inhibited osteogenic differentiation (calcium deposition, alkaline phosphatase activity, and marker gene expression) providing further support for the close coupling of osteodifferentiation and cell death. Metabolic collapse appears to be central in the hydrogen peroxide-PARP1-p38 pathway as silencing PARP1 or inhibition of p38 prevented differentiation-associated loss of cellular NAD, inhibition of mitochondrial respiration, and glycolytic activity. We also provide evidence that endogenous hydrogen peroxide produced by the differentiating cells is sufficient to cause detectable DNA breakage. Moreover, p38 translocates from the cytoplasm to the nucleus where it interacts and colocalizes with PARP1 as detected by immunoprecipitation and immunofluorescence, respectively. In summary, hydrogen peroxide-induced PARP1 activation leads to p38 activation and this pathway is required both for the successful completion of the differentiation process and for the associated cell death.

Original languageEnglish
Pages (from-to)69-79
Number of pages11
JournalFree Radical Biology and Medicine
Volume76
DOIs
Publication statusPublished - 2014

Fingerprint

Poly(ADP-ribose) Polymerases
Cell death
Adenosine Diphosphate
Cell Death
Hydrogen Peroxide
Chemical activation
Poly (ADP-Ribose) Polymerase-1
DNA Breaks
DNA
p38 Mitogen-Activated Protein Kinases
Stem cells
Mesenchymal Stromal Cells
Immunoprecipitation
Gene expression
NAD
Catalase
Fluorescent Antibody Technique
Alkaline Phosphatase
Respiration
Cytoplasm

Keywords

  • Cell death
  • Hydrogen peroxide
  • Osteogenic differentiation
  • p38 MAP kinase
  • PARP1
  • Poly(ADP-ribosyl)ation

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)
  • Medicine(all)

Cite this

The role of p38 signaling and poly(ADP-ribosyl)ation-induced metabolic collapse in the osteogenic differentiation-coupled cell death pathway. / Robaszkiewicz, Agnieszka; Valkó, Zsuzsanna; Kovács, Katalin; Hegedus, Csaba; Bakondi, Edina; Bai, P.; Virag, L.

In: Free Radical Biology and Medicine, Vol. 76, 2014, p. 69-79.

Research output: Contribution to journalArticle

Robaszkiewicz, Agnieszka ; Valkó, Zsuzsanna ; Kovács, Katalin ; Hegedus, Csaba ; Bakondi, Edina ; Bai, P. ; Virag, L. / The role of p38 signaling and poly(ADP-ribosyl)ation-induced metabolic collapse in the osteogenic differentiation-coupled cell death pathway. In: Free Radical Biology and Medicine. 2014 ; Vol. 76. pp. 69-79.
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AU - Kovács, Katalin

AU - Hegedus, Csaba

AU - Bakondi, Edina

AU - Bai, P.

AU - Virag, L.

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AB - Osteogenic differentiation is a multistep process regulated by a diverse set of morphogenic and transcription factors. Previously we identified endogenous hydrogen peroxide-induced poly(ADP-ribose) polymerase-1 (PARP1) activation as a mediator of osteodifferentiation and associated cell death. Here we set out to investigate whether or not activation of PARP1 is dependent on DNA breaks and how PARP1 mediates cell death during osteodifferentiation of mesenchymal stem cells and SAOS-2 cells. Here we show that the MAP kinases p38, JNK, and ERK1/2 become activated during the differentiation process. However, only p38 activation depended both on hydrogen peroxide production and on PARP1 activation as the hydrogen peroxide decomposing enzyme catalase, the PARP inhibitor PJ34, and the silencing of PARP1 suppressed p38 activation. Inhibition of p38 suppressed cell death and inhibited osteogenic differentiation (calcium deposition, alkaline phosphatase activity, and marker gene expression) providing further support for the close coupling of osteodifferentiation and cell death. Metabolic collapse appears to be central in the hydrogen peroxide-PARP1-p38 pathway as silencing PARP1 or inhibition of p38 prevented differentiation-associated loss of cellular NAD, inhibition of mitochondrial respiration, and glycolytic activity. We also provide evidence that endogenous hydrogen peroxide produced by the differentiating cells is sufficient to cause detectable DNA breakage. Moreover, p38 translocates from the cytoplasm to the nucleus where it interacts and colocalizes with PARP1 as detected by immunoprecipitation and immunofluorescence, respectively. In summary, hydrogen peroxide-induced PARP1 activation leads to p38 activation and this pathway is required both for the successful completion of the differentiation process and for the associated cell death.

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