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Poly(ADP-ribosyl)ation of p53 contributes to TPEN-induced neuronal apoptosis.

Kim HL, Ra H, Kim KR, Lee JM, Im H, Kim YH - Mol. Cells (2015)

Bottom Line: Poly(ADP-ribosyl)ation of p53 occurred starting 1 h after TPEN treatment.Consistent with this, the induction of downstream proapoptotic proteins PUMA and NOXA was noticeably reduced by chemical inhibitors or genetic deletion of PARP-1.Taken together, these findings indicate that PARP-1 is essential for TPEN-induced neuronal apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Sejong University, Seoul 143-747, Korea.

ABSTRACT
Depletion of intracellular zinc by N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine (TPEN) induces p53-mediated protein synthesis-dependent apoptosis of mouse cortical neurons. Here, we examined the requirement for poly(ADP-ribose) polymerase (PARP)-1 as an upstream regulator of p53 in zinc depletion-induced neuronal apoptosis. First, we found that chemical inhibition or genetic deletion of PARP-1 markedly attenuated TPEN-induced apoptosis of cultured mouse cortical neurons. Poly(ADP-ribosyl)ation of p53 occurred starting 1 h after TPEN treatment. Suggesting the critical role of PARP-1, the TPEN-induced increase of stability and activity of p53 as well as poly(ADP-ribosyl)ation of p53 was almost completely blocked by PARP inhibition. Consistent with this, the induction of downstream proapoptotic proteins PUMA and NOXA was noticeably reduced by chemical inhibitors or genetic deletion of PARP-1. TPEN-induced cytochrome C release into the cytosol and caspase-3 activation were also blocked by inhibition of PARP-1. Taken together, these findings indicate that PARP-1 is essential for TPEN-induced neuronal apoptosis.

No MeSH data available.


Related in: MedlinePlus

PARP-1 as an upstream regulator of TPEN-induced pro-apoptotic proteins. (A) RTPCR (left) and western blot analysis (right) of PUMA, NOXA, and Bax expression. RNA or protein samples were prepared from neuron cultures after 6-h exposure to sham wash (CTRL) or TPEN with or without NAM or AB. (B) Western blot analysis of PUMA, NOXA, and Bax expression. Protein samples were prepared from PARP-1+/+ or PARP-1−/− mouse cortical neuronal cultures after 6-h exposure to sham wash (CTRL) or TPEN. Inhibition of PARP-1 by chemical inhibitors or genetic deletion almost completely reduced the induction of proapoptotic proteins by TPEN.
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f3-molce-38-4-312: PARP-1 as an upstream regulator of TPEN-induced pro-apoptotic proteins. (A) RTPCR (left) and western blot analysis (right) of PUMA, NOXA, and Bax expression. RNA or protein samples were prepared from neuron cultures after 6-h exposure to sham wash (CTRL) or TPEN with or without NAM or AB. (B) Western blot analysis of PUMA, NOXA, and Bax expression. Protein samples were prepared from PARP-1+/+ or PARP-1−/− mouse cortical neuronal cultures after 6-h exposure to sham wash (CTRL) or TPEN. Inhibition of PARP-1 by chemical inhibitors or genetic deletion almost completely reduced the induction of proapoptotic proteins by TPEN.

Mentions: We previously showed that Bcl-2 homology domain 3 (BH3)-only proteins, such as PUMA and NOXA, are induced by TPEN in a p53-dependent manner (Ra et al., 2009). Here, we found that increases in BH3-only pro-apoptotic proteins were mediated by PARP-1. Induction of PUMA and NOXA expression by TPEN was markedly reduced by chemical inhibitors of PARP (Fig. 3A). In PARP-1−/− neuronal cultures, the protein expression level of PUMA or NOXA was not increased by TPEN (Fig. 3B), indicating mediation by PARP-1. Bax, expression levels were not affected by TPEN in either PARP-1+/+ or PARP-1−/− cortical neuron cultures (Fig. 3B). Consistently, the chemical inhibition or genetic deletion of PARP-1 had no effect on Bax expression levels (Fig. 3B).


Poly(ADP-ribosyl)ation of p53 contributes to TPEN-induced neuronal apoptosis.

Kim HL, Ra H, Kim KR, Lee JM, Im H, Kim YH - Mol. Cells (2015)

PARP-1 as an upstream regulator of TPEN-induced pro-apoptotic proteins. (A) RTPCR (left) and western blot analysis (right) of PUMA, NOXA, and Bax expression. RNA or protein samples were prepared from neuron cultures after 6-h exposure to sham wash (CTRL) or TPEN with or without NAM or AB. (B) Western blot analysis of PUMA, NOXA, and Bax expression. Protein samples were prepared from PARP-1+/+ or PARP-1−/− mouse cortical neuronal cultures after 6-h exposure to sham wash (CTRL) or TPEN. Inhibition of PARP-1 by chemical inhibitors or genetic deletion almost completely reduced the induction of proapoptotic proteins by TPEN.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400305&req=5

f3-molce-38-4-312: PARP-1 as an upstream regulator of TPEN-induced pro-apoptotic proteins. (A) RTPCR (left) and western blot analysis (right) of PUMA, NOXA, and Bax expression. RNA or protein samples were prepared from neuron cultures after 6-h exposure to sham wash (CTRL) or TPEN with or without NAM or AB. (B) Western blot analysis of PUMA, NOXA, and Bax expression. Protein samples were prepared from PARP-1+/+ or PARP-1−/− mouse cortical neuronal cultures after 6-h exposure to sham wash (CTRL) or TPEN. Inhibition of PARP-1 by chemical inhibitors or genetic deletion almost completely reduced the induction of proapoptotic proteins by TPEN.
Mentions: We previously showed that Bcl-2 homology domain 3 (BH3)-only proteins, such as PUMA and NOXA, are induced by TPEN in a p53-dependent manner (Ra et al., 2009). Here, we found that increases in BH3-only pro-apoptotic proteins were mediated by PARP-1. Induction of PUMA and NOXA expression by TPEN was markedly reduced by chemical inhibitors of PARP (Fig. 3A). In PARP-1−/− neuronal cultures, the protein expression level of PUMA or NOXA was not increased by TPEN (Fig. 3B), indicating mediation by PARP-1. Bax, expression levels were not affected by TPEN in either PARP-1+/+ or PARP-1−/− cortical neuron cultures (Fig. 3B). Consistently, the chemical inhibition or genetic deletion of PARP-1 had no effect on Bax expression levels (Fig. 3B).

Bottom Line: Poly(ADP-ribosyl)ation of p53 occurred starting 1 h after TPEN treatment.Consistent with this, the induction of downstream proapoptotic proteins PUMA and NOXA was noticeably reduced by chemical inhibitors or genetic deletion of PARP-1.Taken together, these findings indicate that PARP-1 is essential for TPEN-induced neuronal apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Sejong University, Seoul 143-747, Korea.

ABSTRACT
Depletion of intracellular zinc by N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine (TPEN) induces p53-mediated protein synthesis-dependent apoptosis of mouse cortical neurons. Here, we examined the requirement for poly(ADP-ribose) polymerase (PARP)-1 as an upstream regulator of p53 in zinc depletion-induced neuronal apoptosis. First, we found that chemical inhibition or genetic deletion of PARP-1 markedly attenuated TPEN-induced apoptosis of cultured mouse cortical neurons. Poly(ADP-ribosyl)ation of p53 occurred starting 1 h after TPEN treatment. Suggesting the critical role of PARP-1, the TPEN-induced increase of stability and activity of p53 as well as poly(ADP-ribosyl)ation of p53 was almost completely blocked by PARP inhibition. Consistent with this, the induction of downstream proapoptotic proteins PUMA and NOXA was noticeably reduced by chemical inhibitors or genetic deletion of PARP-1. TPEN-induced cytochrome C release into the cytosol and caspase-3 activation were also blocked by inhibition of PARP-1. Taken together, these findings indicate that PARP-1 is essential for TPEN-induced neuronal apoptosis.

No MeSH data available.


Related in: MedlinePlus