Functional analysis of the human mitochondrial proteome

While we understand many aspects of mitochondrial function, we still do not fully appreciate the roles played by many individual proteins present in the organelle. Of the ~1300 proteins in human mitochondria, about 30% have no known function while another 30% have been assigned functions based on sequence similarity. We are coupling gene-editing techniques (TALENs and CRISPRs) to generate gene knockouts in cultured human cells and investigating how a protein's loss impacts mitochondria and the cell. This work is being coupled with state-of-the-art proteomic and bioinformatic approaches to determine how the cell changes its protein make-up to adjust to these changes. In our approach to functionalise the human mitochondrial proteome we are making new and important discoveries into fundamental aspects related to mitochondrial protein function.

 

Recent publications:

Stroud DA, Surgenor EE, Formosa LE, Reljic B, Frazier AE, Dibley MG, Osellame LD, Stait T, Beilharz TH, Thorburn DR, Salim A &  Ryan MT. Accessory subunits are integral for assembly and function of human mitochondrial complex I. Nature (14 September 2016) | doi:10.1038/nature19754 Pubmed http://rdcu.be/kkQg

Osellame LD, Singh AP, Stroud DA, Palmer CS, Stojanovski D, Ramachandran R, Ryan MT. Cooperative and independent roles of Drp1 adaptors Mff and MiD49/51 in mitochondrial fission. J Cell Sci. 2016 Apr 12. pii: jcs.185165. [Epub ahead of print] Pubmed

Stroud DA, Maher MJ, Lindau C, Vögtle FN, Frazier AE, Surgenor E, Mountford H, Singh AP, Bonas M, Oeljeklaus S, Warscheid B, Meisinger C, Thorburn DR, Ryan MT. COA6 is a mitochondrial complex IV assembly factor critical for biogenesis of mtDNA-encoded COX2. Hum Mol Genet. (2015) 24(19):5404-15. Pubmed

Formosa LE, Mimaki M, Frazier AE, McKenzie M, Stait TL, Thorburn DR, Stroud DA, Ryan MT. Characterization of mitochondrial FOXRED1 in the assembly of respiratory chain complex I. Hum Mol Genet. (2015) 24(10):2952-65 Pubmed

Stroud DA, Formosa LE, Wijeyeratne XW, Nguyen TN, Ryan MT. Gene knockout using transcription activator-like effector nucleases (TALENs) reveals that human NDUFA9 protein is essential for stabilizing the junction between membrane and matrix arms of complex I. J Biol Chem. 2013288(3):1685-90 Pubmed