Rat Humanin is encoded and translated in mitochondria and is localized to the mitochondrial compartment where it regulates ROS production

•Evidence for the putative mitochondrial origin of the cytoprotective Humanin (HN) peptide has been lacking.•The rat HN (Rattin; rHN) homologue is derived from the mitochondrial genome and is translated in mitochondria.•rHN localizes to the mitochondrial compartment and reduces hydrogen peroxide production.•We establish proof-of-principle for a bioactive peptide encoded from an open reading frame in the rat mitochondrial 16S rRNA.

Evidence for the putative mitochondrial origin of the Humanin (HN) peptide has been lacking, although its cytoprotective activity has been demonstrated in a variety of organismal and cellular systems. We sought to establish proof-of-principle for a mitochondria-derived peptide (MDP) in a rat-derived cellular system as the rat HN sequence is predicted to lack nuclear insertions of mitochondrial origin (NUMT). We found that the rat HN (Rattin; rHN) homologue is derived from the mitochondrial genome as evidenced by decreased production in Rho-0 cells, and that peptide translation occurs in the mitochondria as it is unaffected by cycloheximide. Rat HN localizes to the mitochondria in cellular subfractionation and immunohistochemical studies. Addition of a HN analogue to isolated mitochondria from rat INS-1 beta cells reduced hydrogen peroxide production by 55%. In summary, a locally bioactive peptide is derived and translated from an open reading frame (ORF) within rat mitochondrial DNA encoding 16S rRNA.