Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling

Pankaj Chaudhary, Rajendra Sharma, Abha Sharma, Rit Vatsyayan, Sushma Yadav, Sharad S. Singhal, Navin Rauniyar, Laszlo Prokai, Sanjay Awasthi, Yogesh C. Awasthi

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

In recent years, 4-hydroxy-2-nonenal (4-HNE) has emerged as an important second messenger in cell cycle signaling. Here, we demonstrate that 4-HNE induces signaling for apoptosis via both the Fas-mediated extrinsic and the p53-mediated intrinsic pathways in HepG2 cells. 4-HNE induces a Fas-mediated DISC independent apoptosis pathway by activating ASK1, JNK, and caspase-3. Parallel treatment of 4-HNE to HepG2 cells also induces apoptosis by the p53 pathway through activation of Bax, p21, JNK, and caspase-3. Exposure of HepG2 cells to 4-HNE leads to the activation of both Fas and Daxx, promotes the export of Daxx from the nucleus to cytoplasm, and facilitates Fas-Daxx binding. Depletion of Daxx by siRNA results in the potentiation of apoptosis, indicating that Fas-Daxx binding in fact is inhibitory to Fas-mediated apoptosis in cells. 4-HNE-induced translocation of Daxx is also accompanied by the activation and nuclear accumulation of HSF1 and up-regulation of heat shock protein Hsp70. All these effects of 4-HNE in cells can be attenuated by ectopic expression of hGSTA4-4, the isozyme of glutathione S-transferase with high activity for 4-HNE. Through immunoprecipitation and liquid chromatography-tandem mass spectrometry, we have demonstrated the covalent binding of 4-HNE to Daxx. We also demonstrate that 4-HNE modification induces phosphorylation of Daxx at Ser668 and Ser671 to facilitate its cytoplasmic export. These results indicate that while 4-HNE exhibits toxicity through several mechanisms, in parallel it evokes signaling for defense mechanisms to self-regulate its toxicity and can simultaneously affect multiple signaling pathways through its interactions with membrane receptors and transcription factors/repressors.

Original languageEnglish
Pages (from-to)6263-6275
Number of pages13
JournalBiochemistry
Volume49
Issue number29
DOIs
StatePublished - 27 Jul 2010

Fingerprint

Apoptosis
Hep G2 Cells
Chemical activation
Caspase 3
Toxicity
4-hydroxy-2-nonenal
Phosphorylation
Liquid chromatography
Second Messenger Systems
Heat-Shock Proteins
Tandem Mass Spectrometry
Glutathione Transferase
Immunoprecipitation
Liquid Chromatography
Small Interfering RNA
Isoenzymes
Mass spectrometry
Cell Cycle
Cytoplasm
Transcription Factors

Cite this

Chaudhary, P., Sharma, R., Sharma, A., Vatsyayan, R., Yadav, S., Singhal, S. S., ... Awasthi, Y. C. (2010). Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling. Biochemistry, 49(29), 6263-6275. https://doi.org/10.1021/bi100517x
Chaudhary, Pankaj ; Sharma, Rajendra ; Sharma, Abha ; Vatsyayan, Rit ; Yadav, Sushma ; Singhal, Sharad S. ; Rauniyar, Navin ; Prokai, Laszlo ; Awasthi, Sanjay ; Awasthi, Yogesh C. / Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling. In: Biochemistry. 2010 ; Vol. 49, No. 29. pp. 6263-6275.
@article{e2337e1bb084476784228a23da573041,
title = "Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling",
abstract = "In recent years, 4-hydroxy-2-nonenal (4-HNE) has emerged as an important second messenger in cell cycle signaling. Here, we demonstrate that 4-HNE induces signaling for apoptosis via both the Fas-mediated extrinsic and the p53-mediated intrinsic pathways in HepG2 cells. 4-HNE induces a Fas-mediated DISC independent apoptosis pathway by activating ASK1, JNK, and caspase-3. Parallel treatment of 4-HNE to HepG2 cells also induces apoptosis by the p53 pathway through activation of Bax, p21, JNK, and caspase-3. Exposure of HepG2 cells to 4-HNE leads to the activation of both Fas and Daxx, promotes the export of Daxx from the nucleus to cytoplasm, and facilitates Fas-Daxx binding. Depletion of Daxx by siRNA results in the potentiation of apoptosis, indicating that Fas-Daxx binding in fact is inhibitory to Fas-mediated apoptosis in cells. 4-HNE-induced translocation of Daxx is also accompanied by the activation and nuclear accumulation of HSF1 and up-regulation of heat shock protein Hsp70. All these effects of 4-HNE in cells can be attenuated by ectopic expression of hGSTA4-4, the isozyme of glutathione S-transferase with high activity for 4-HNE. Through immunoprecipitation and liquid chromatography-tandem mass spectrometry, we have demonstrated the covalent binding of 4-HNE to Daxx. We also demonstrate that 4-HNE modification induces phosphorylation of Daxx at Ser668 and Ser671 to facilitate its cytoplasmic export. These results indicate that while 4-HNE exhibits toxicity through several mechanisms, in parallel it evokes signaling for defense mechanisms to self-regulate its toxicity and can simultaneously affect multiple signaling pathways through its interactions with membrane receptors and transcription factors/repressors.",
author = "Pankaj Chaudhary and Rajendra Sharma and Abha Sharma and Rit Vatsyayan and Sushma Yadav and Singhal, {Sharad S.} and Navin Rauniyar and Laszlo Prokai and Sanjay Awasthi and Awasthi, {Yogesh C.}",
year = "2010",
month = "7",
day = "27",
doi = "10.1021/bi100517x",
language = "English",
volume = "49",
pages = "6263--6275",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "29",

}

Chaudhary, P, Sharma, R, Sharma, A, Vatsyayan, R, Yadav, S, Singhal, SS, Rauniyar, N, Prokai, L, Awasthi, S & Awasthi, YC 2010, 'Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling', Biochemistry, vol. 49, no. 29, pp. 6263-6275. https://doi.org/10.1021/bi100517x

Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling. / Chaudhary, Pankaj; Sharma, Rajendra; Sharma, Abha; Vatsyayan, Rit; Yadav, Sushma; Singhal, Sharad S.; Rauniyar, Navin; Prokai, Laszlo; Awasthi, Sanjay; Awasthi, Yogesh C.

In: Biochemistry, Vol. 49, No. 29, 27.07.2010, p. 6263-6275.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling

AU - Chaudhary, Pankaj

AU - Sharma, Rajendra

AU - Sharma, Abha

AU - Vatsyayan, Rit

AU - Yadav, Sushma

AU - Singhal, Sharad S.

AU - Rauniyar, Navin

AU - Prokai, Laszlo

AU - Awasthi, Sanjay

AU - Awasthi, Yogesh C.

PY - 2010/7/27

Y1 - 2010/7/27

N2 - In recent years, 4-hydroxy-2-nonenal (4-HNE) has emerged as an important second messenger in cell cycle signaling. Here, we demonstrate that 4-HNE induces signaling for apoptosis via both the Fas-mediated extrinsic and the p53-mediated intrinsic pathways in HepG2 cells. 4-HNE induces a Fas-mediated DISC independent apoptosis pathway by activating ASK1, JNK, and caspase-3. Parallel treatment of 4-HNE to HepG2 cells also induces apoptosis by the p53 pathway through activation of Bax, p21, JNK, and caspase-3. Exposure of HepG2 cells to 4-HNE leads to the activation of both Fas and Daxx, promotes the export of Daxx from the nucleus to cytoplasm, and facilitates Fas-Daxx binding. Depletion of Daxx by siRNA results in the potentiation of apoptosis, indicating that Fas-Daxx binding in fact is inhibitory to Fas-mediated apoptosis in cells. 4-HNE-induced translocation of Daxx is also accompanied by the activation and nuclear accumulation of HSF1 and up-regulation of heat shock protein Hsp70. All these effects of 4-HNE in cells can be attenuated by ectopic expression of hGSTA4-4, the isozyme of glutathione S-transferase with high activity for 4-HNE. Through immunoprecipitation and liquid chromatography-tandem mass spectrometry, we have demonstrated the covalent binding of 4-HNE to Daxx. We also demonstrate that 4-HNE modification induces phosphorylation of Daxx at Ser668 and Ser671 to facilitate its cytoplasmic export. These results indicate that while 4-HNE exhibits toxicity through several mechanisms, in parallel it evokes signaling for defense mechanisms to self-regulate its toxicity and can simultaneously affect multiple signaling pathways through its interactions with membrane receptors and transcription factors/repressors.

AB - In recent years, 4-hydroxy-2-nonenal (4-HNE) has emerged as an important second messenger in cell cycle signaling. Here, we demonstrate that 4-HNE induces signaling for apoptosis via both the Fas-mediated extrinsic and the p53-mediated intrinsic pathways in HepG2 cells. 4-HNE induces a Fas-mediated DISC independent apoptosis pathway by activating ASK1, JNK, and caspase-3. Parallel treatment of 4-HNE to HepG2 cells also induces apoptosis by the p53 pathway through activation of Bax, p21, JNK, and caspase-3. Exposure of HepG2 cells to 4-HNE leads to the activation of both Fas and Daxx, promotes the export of Daxx from the nucleus to cytoplasm, and facilitates Fas-Daxx binding. Depletion of Daxx by siRNA results in the potentiation of apoptosis, indicating that Fas-Daxx binding in fact is inhibitory to Fas-mediated apoptosis in cells. 4-HNE-induced translocation of Daxx is also accompanied by the activation and nuclear accumulation of HSF1 and up-regulation of heat shock protein Hsp70. All these effects of 4-HNE in cells can be attenuated by ectopic expression of hGSTA4-4, the isozyme of glutathione S-transferase with high activity for 4-HNE. Through immunoprecipitation and liquid chromatography-tandem mass spectrometry, we have demonstrated the covalent binding of 4-HNE to Daxx. We also demonstrate that 4-HNE modification induces phosphorylation of Daxx at Ser668 and Ser671 to facilitate its cytoplasmic export. These results indicate that while 4-HNE exhibits toxicity through several mechanisms, in parallel it evokes signaling for defense mechanisms to self-regulate its toxicity and can simultaneously affect multiple signaling pathways through its interactions with membrane receptors and transcription factors/repressors.

UR - http://www.scopus.com/inward/record.url?scp=77954824524&partnerID=8YFLogxK

U2 - 10.1021/bi100517x

DO - 10.1021/bi100517x

M3 - Article

C2 - 20565132

AN - SCOPUS:77954824524

VL - 49

SP - 6263

EP - 6275

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 29

ER -

Chaudhary P, Sharma R, Sharma A, Vatsyayan R, Yadav S, Singhal SS et al. Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling. Biochemistry. 2010 Jul 27;49(29):6263-6275. https://doi.org/10.1021/bi100517x