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Expression of FADD and cFLIPL balances mitochondrial integrity and redox signaling to substantiate apoptotic cell death

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Abstract

FADD and cFLIP both are pivotal components of death receptor signaling. The cellular signaling of apoptosis accomplished with death receptors and mitochondria follows independent pathways for cell death. FADD and cFLIP both have an important role in the regulation of apoptotic and non-apoptotic functions. Dysregulated expression of FADD and cFLIP is associated with resistance to apoptosis in cancer cells. Mitochondria are known to play critical role in maintaining cellular respiration and homeostasis in the cells as well as transduces various signals to determine the fate of cell death. However, involvement of FADD and cFLIP in regulation of mitochondrial integrity and programmed cell death signaling to define the fate of cells remains elusive. In the present study, we explored that, induced expression of FADD challenges the mitochondrial integrity and pulverizes the membrane potential by altering the expression of Bcl-2 and cytochrome c. In contrast, mutant of FADD was unable to affect the mitochondrial integrity. Interestingly, expression of FADD and cFLIP helps to balance redox potential by regulating the anti-oxidant levels. Further, we noticed that, knockdown of cFLIPL and induced expression of FADD rapidly accumulate intracellular ROS accompanied by JNK1 activation to substantiate apoptosis. Notably, the ectopic expression of cFLIPL resists the sensitivity of cancer cells against apoptosis inducers Etoposide and HA14-1. Altogether, our findings suggest that FADD and cFLIPL are important modulators of mitochondrial-associated apoptosis apart from the death receptor signaling.

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Abbreviations

cFLIP:

Cellular FLICE-like inhibitory protein

cFLIP KDL :

Knockdown of cFLIP large isoform

DR:

Death receptor

FADD:

Fas-associated death domain

JNK1:

c-jun-N-terminal kinase-1

MMP:

Mitochondrial membrane potential

MOM:

Mitochondrial outer membrane

NAC:

N-acetyl cysteine

ROS:

Reactive oxygen species

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Acknowledgments

We are thankful to Dr. Andrew Thorburn, (University of Colorado, USA), Dr. Eyal Gottlieb (Cancer Research UK, Beatson Institute), Dr. R. C. Bleackley (University of Alberta, Canada), Dr. Douglas Green (St. Jude Children’s Research Hospital, USA), Dr. Penelope Duerksen-Hughes (Loma Linda University, USA), Prof. Tamotsu Yoshimori (Osaka University, Japan), and Prof. Sabine Adam (Universitätsklinikum Schleswig–Holstein, Germany) for sharing plasmid constructs. This work was supported by the research grants scheme Cancer Research Pilot Project received from the Department of Biotechnology, Ministry of Science & Technology, Government of India to CP. The Indian Council of Medical Research, Govt. of India is acknowledged for providing Senior Research Fellowship to KR.

Author contributions

K. R. performed all the experiments. K. R and C. P. designed the experiments, analyzed the data, and wrote the manuscript.

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  1. Department of Cell Biology, School of Biological Sciences and Biotechnology, Indian Institute of Advanced Research, Koba Institutional Area, Gandhinagar, Gujarat, 382007, India

    Kishu Ranjan & Chandramani Pathak

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  1. Kishu Ranjan
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Correspondence to Chandramani Pathak.

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Ranjan, K., Pathak, C. Expression of FADD and cFLIPL balances mitochondrial integrity and redox signaling to substantiate apoptotic cell death. Mol Cell Biochem 422, 135–150 (2016). https://doi.org/10.1007/s11010-016-2813-z

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