Molecular insights on cytochrome c and nucleotide regulation of apoptosome function and its implication in cancer.
Alanine
/ chemistry
Amino Acid Substitution
Apoptosomes
/ chemistry
Apoptotic Protease-Activating Factor 1
/ chemistry
Case-Control Studies
Cell Transformation, Neoplastic
/ genetics
Cells, Cultured
Cytochromes c
/ chemistry
Female
Humans
Lysine
/ chemistry
Male
Models, Molecular
Molecular Docking Simulation
Mutant Proteins
/ chemistry
Neoplasms
/ genetics
Nucleotides
/ chemistry
PC-3 Cells
Protein Binding
/ genetics
Protein Interaction Mapping
Protein Multimerization
/ genetics
Signal Transduction
/ genetics
Apoptosome
Caspase activation
Cytochrome c
Mitochondria
Nucleotides
Tumorigenesis
Journal
Biochimica et biophysica acta. Molecular cell research
ISSN: 1879-2596
Titre abrégé: Biochim Biophys Acta Mol Cell Res
Pays: Netherlands
ID NLM: 101731731
Informations de publication
Date de publication:
01 2020
01 2020
Historique:
received:
28
03
2019
revised:
14
10
2019
accepted:
22
10
2019
pubmed:
5
11
2019
medline:
21
4
2020
entrez:
4
11
2019
Statut:
ppublish
Résumé
Cytochrome c (Cyt c) released from mitochondria interacts with Apaf-1 to form the heptameric apoptosome, which initiates the caspase cascade to execute apoptosis. Although lysine residue at 72 (K72) of Cyt c plays an important role in the Cyt c-Apaf-1 interaction, the underlying mechanism of interaction between Cyt c and Apaf-1 is still not clearly defined. Here we identified multiple lysine residues including K72, which are also known to interact with ATP, to play a key role in Cyt c-Apaf-1 interaction. Mutation of these lysine residues abrogates the apoptosome formation causing inhibition of caspase activation. Using in-silico molecular docking, we have identified Cyt c-binding interface on Apaf-1. Although mutant Cyt c shows higher affinity for Apaf-1, the presence of Cyt c-WT restores the apoptosome activity. ATP addition modulates only mutant Cyt c binding to Apaf-1 but not WT Cyt c binding to Apaf-1. Using TCGA and cBioPortal, we identified multiple mutations in both Apaf-1 and Cyt c that are predicted to interfere with apoptosome assembly. We also demonstrate that transcript levels of various enzymes involved with dATP or ATP synthesis are increased in various cancers. Silencing of nucleotide metabolizing enzymes such as ribonucleotide reductase subunit M1 (RRM1) and ATP-producing glycolytic enzymes PKM2 attenuated ATP production and enhanced caspase activation. These findings suggest important role for lysine residues of Cyt c and nucleotides in the regulation of apoptosome-dependent apoptotic cell death as well as demonstrate how these mutations and nucleotides may have a pivotal role in human diseases such as cancer.
Identifiants
pubmed: 31678591
pii: S0167-4889(19)30181-8
doi: 10.1016/j.bbamcr.2019.118573
pmc: PMC7733678
mid: NIHMS1649871
pii:
doi:
Substances chimiques
APAF1 protein, human
0
Apoptosomes
0
Apoptotic Protease-Activating Factor 1
0
Mutant Proteins
0
Nucleotides
0
Cytochromes c
9007-43-6
Lysine
K3Z4F929H6
Alanine
OF5P57N2ZX
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
118573Subventions
Organisme : NCI NIH HHS
ID : K01 CA123142
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA160685
Pays : United States
Informations de copyright
Copyright © 2019 Elsevier B.V. All rights reserved.
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