Human mitochondrial RNA decay mediated by PNPase-hSuv3 complex takes place in distinct foci

• Published in: Nucleic acids research Vol. 41; no. 2; pp. 1223 - 1240
• Main Authors: Borowski, Lukasz S, Dziembowski, Andrzej, Hejnowicz, Monika S, Stepien, Piotr P, Szczesny, Roman J
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-01-2013
• Subjects: Adenosine Triphosphate - metabolism; Cell Growth Processes; Cell Line; Cytoplasm; DEAD-box RNA Helicases - analysis; DEAD-box RNA Helicases - metabolism; DNA helicase; DNA, Mitochondrial - analysis; Endoribonucleases - metabolism; Fluorescence; Gene Silencing; Humans; imaging; Microscopy; Mitochondria; Mitochondria - enzymology; Mitochondrial Proteins - metabolism; Multienzyme Complexes - metabolism; Nucleoids; polynucleotide phosphorylase; Polyribonucleotide Nucleotidyltransferase - analysis; Polyribonucleotide Nucleotidyltransferase - genetics; Polyribonucleotide Nucleotidyltransferase - metabolism; Ribonuclease; RNA; RNA - analysis; RNA - metabolism; RNA Helicases - metabolism; RNA Stability
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gks1130

RNA decay is usually mediated by protein complexes and can occur in specific foci such as P-bodies in the cytoplasm of eukaryotes. In human mitochondria nothing is known about the spatial organization of the RNA decay machinery, and the ribonuclease responsible for RNA degradation has not been identified. We demonstrate that silencing of human polynucleotide phosphorylase (PNPase) causes accumulation of RNA decay intermediates and increases the half-life of mitochondrial transcripts. A combination of fluorescence lifetime imaging microscopy with Förster resonance energy transfer and bimolecular fluorescence complementation (BiFC) experiments prove that PNPase and hSuv3 helicase (Suv3, hSuv3p and SUPV3L1) form the RNA-degrading complex in vivo in human mitochondria. This complex, referred to as the degradosome, is formed only in specific foci (named D-foci), which co-localize with mitochondrial RNA and nucleoids. Notably, interaction between PNPase and hSuv3 is essential for efficient mitochondrial RNA degradation. This provides indirect evidence that degradosome-dependent mitochondrial RNA decay takes place in foci.