RT Journal Article SR Electronic T1 Ancestral sequence reconstruction of Mic60 reveals a residue signature supporting respiration in yeast JF bioRxiv FD Cold Spring Harbor Laboratory SP 2024.04.26.591372 DO 10.1101/2024.04.26.591372 A1 Benning, Friederike M. C. A1 Bell, Tristan A. A1 Nguyen, Tran H. A1 Syau, Della A1 Connell, Louise B. A1 daCosta, Corrie J. B. A1 Chao, Luke H. YR 2024 UL http://biorxiv.org/content/early/2024/04/29/2024.04.26.591372.abstract AB In eukaryotes, the essential process of cellular respiration takes place in the cristae of mitochondria. The protein Mic60 is known to stabilize crista junctions; however, how the C-terminal Mitofilin domain of Mic60 mediates cristae-supported respiration remains elusive. Here, we used ancestral sequence reconstruction to generate Mitofilin ancestors up to and including the last opisthokont common ancestor (LOCA). We found that yeast-lineage derived Mitofilin ancestors as far back as the LOCA rescue respiration. By comparing Mitofilin ancestors with different respiratory phenotypes, we identify four residues that explain the difference between respiration functional yeast- and non-functional animal-derived common Mitofilin ancestors. Our results imply that Mitofilin-supported respiration in yeast stems from a conserved mechanism, and provide a foundation for investigating the divergence of candidate crista junction interactions present during the emergence of eukaryotes.Competing Interest StatementThe authors have declared no competing interest.