title: Transformations of the macromolecular landscape at mitochondria during DNA-damage-induced apoptotic cell death.
abstract: Apoptosis is a dynamic process regulated by mitochondrion critical for cellular respiration and survival. Execution of apoptosis is mediated by multiple protein signaling events at mitochondria. Initiation and progression of apoptosis require numerous apoptogenic factors that are either released from or sequestered in mitochondria, which may transform the biomolecular makeup of the organelle. In this communication, using Raman microspectroscopy, we demonstrate that transformation in biomolecular composition of mitochondrion may be used as apoptosis marker in an individual cell. For the first time, we show that significant changes occur in the concentrations of RNA, DNA, protein, and lipid constituents of mitochondria during apoptosis. The structural analysis of proteins on mitochondria demonstrated a decrease in α-helix secondary structure content, and an increase in the levels of random coils and β-sheets on mitochondria. This may represent an additional hallmark of apoptosis. Strikingly, we observed nearly identical changes in macromolecular content of mitochondria both in the presence and absence of a key proapoptotic protein, Bax (Bcl-2-associated X protein). Increased DNA level in mitochondria corresponded with higher mitochondrial DNA (mtDNA), cellular reactive oxygen species (ROS), and mitochondrial ROS production. Upregulation of polymerase-γ (POLG), mitochondrial helicase Twinkle, and mitochondrial transcription factor A (Tfam) in response to DNA damage correlated with increased mtDNA and RNA synthesis. Elevated activity of oxidative phosphorylation complexes supports functional mitochondrial respiration during apoptosis. Thus, we define previously unknown dynamic correlation of macromolecular structure of mitochondria and apoptosis progression in the presence and absence of Bax protein. These findings open up a new approach for monitoring physiological status of cells by non invasive single-cell method.
authors.affiliation: Department of Pharmacology and Therapeutics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY, USA.
authors.forename: N
authors.initials: N
authors.lastname: Yadav
authors.affiliation: Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY, USA.
authors.forename: A
authors.initials: A
authors.lastname: Pliss
authors.affiliation: Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY, USA.
authors.forename: A
authors.initials: A
authors.lastname: Kuzmin
authors.affiliation: Department of Pharmacology and Therapeutics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY, USA.
authors.forename: P
authors.initials: P
authors.lastname: Rapali
authors.affiliation: 1] Department of Pharmacology and Therapeutics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY, USA [2] Gastrointestinal Division, Sir Run Run Shaw Hospital, Zhejiang University Medical School, Hangzhou, China.
authors.forename: L
authors.initials: L
authors.lastname: Sun
authors.affiliation: Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY, USA.
authors.forename: P
authors.initials: P
authors.lastname: Prasad
authors.affiliation: Department of Pharmacology and Therapeutics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY, USA.
authors.forename: D
authors.initials: D
authors.lastname: Chandra
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D004249:DNA Damage; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry / Q000378:metabolism*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D004272:DNA, Mitochondrial; D046911:Macromolecular Substances; D051028:bcl-2-Associated X Protein