ChChd3, an Inner Mitochondrial Membrane Protein, Is Essential for Maintaining Crista Integrity and Mitochondrial Function

• Published in: The Journal of biological chemistry Vol. 286; no. 4; pp. 2918 - 2932
• Main Authors: Darshi, Manjula, Mendiola, Vincent L., Mackey, Mason R., Murphy, Anne N., Koller, Antonius, Perkins, Guy A., Ellisman, Mark H., Taylor, Susan S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28.01.2011; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Bioenergetics; ChChd3; Confocal Microscopy; Crista Junctions; Crista Morphology; Electron Microscopy (EM); HeLa Cells; Humans; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Mitochondria; Mitochondria, Liver - genetics; Mitochondria, Liver - metabolism; Mitochondrial Dynamics; Mitochondrial Membranes - metabolism; Mitochondrial Proteins - genetics; Mitochondrial Proteins - metabolism; Multiprotein Complexes - genetics; Multiprotein Complexes - metabolism; Muscle Proteins - genetics; Muscle Proteins - metabolism; Protein Import; Protein-Protein Interactions; Mitochondrial Dynamics; Mitochondria; Bioenergetics; ChChd3; Crista Junctions; Confocal Microscopy; Protein Import; Electron Microscopy (EM); Protein-Protein Interactions; Crista Morphology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.171975

The mitochondrial inner membrane (IM) serves as the site for ATP production by hosting the oxidative phosphorylation complex machinery most notably on the crista membranes. Disruption of the crista structure has been implicated in a variety of cardiovascular and neurodegenerative diseases. Here, we characterize ChChd3, a previously identified PKA substrate of unknown function (Schauble, S., King, C. C., Darshi, M., Koller, A., Shah, K., and Taylor, S. S. (2007) J. Biol. Chem. 282, 14952–14959), and show that it is essential for maintaining crista integrity and mitochondrial function. In the mitochondria, ChChd3 is a peripheral protein of the IM facing the intermembrane space. RNAi knockdown of ChChd3 in HeLa cells resulted in fragmented mitochondria, reduced OPA1 protein levels and impaired fusion, and clustering of the mitochondria around the nucleus along with reduced growth rate. Both the oxygen consumption and glycolytic rates were severely restricted. Ultrastructural analysis of these cells revealed aberrant mitochondrial IM structures with fragmented and tubular cristae or loss of cristae, and reduced crista membrane. Additionally, the crista junction opening diameter was reduced to 50% suggesting remodeling of cristae in the absence of ChChd3. Analysis of the ChChd3-binding proteins revealed that ChChd3 interacts with the IM proteins mitofilin and OPA1, which regulate crista morphology, and the outer membrane protein Sam50, which regulates import and assembly of β-barrel proteins on the outer membrane. Knockdown of ChChd3 led to almost complete loss of both mitofilin and Sam50 proteins and alterations in several mitochondrial proteins, suggesting that ChChd3 is a scaffolding protein that stabilizes protein complexes involved in maintaining crista architecture and protein import and is thus essential for maintaining mitochondrial structure and function.