A novel mitochondrial nuclease‐associated protein: A major executor of the programmed nuclear death in Tetrahymena thermophila

Background Information Programmed nuclear death (PND) in the ciliate Tetrahymena is an apoptosis‐like phenomenon that occurs in a restricted space of cytoplasm during conjugation. In the process, only the parental macronucleus is selectively eliminated from the progeny cytoplasm, in conjunction with differentiation of new macronuclei for the next generation. For the last decade, mitochondria have been elucidated to be a crucial executioner like apoptosis: apoptosis‐inducing factor and yet‐unidentified nucleases localised in mitochondria are major factors for PND. Results To identify such nucleases, we performed a DNase assay in a PAGE (SDS‐DNA‐PAGE) using total mitochondrial proteins. Some proteins showed DNase activity, but particularly a 17 kDa protein exhibited the highest and predominant activity. Mass spectrometric analysis revealed a novel mitochondrial nuclease, named TMN1, whose homologue has been discovered only in the ciliate Paramecium tetraurelia , but not in other eukaryotes. Gene disruption of TMN1 led to a drastic reduction of mitochondrial nuclease activity and blocked nuclear degradation during conjugation, but did not affect accumulation of autophagic and lysosomal machinery around the parental macronucleus. Conclusions These observations strongly suggest that the mitochondrial nuclease‐associated protein plays a key role in PND as a major executor. Taking the novel protein specific to ciliates in consideration, Tetrahymena would have diverted a different protein from common apoptotic factors shared in eukaryotes to PND in the course of ciliate evolution.