Novel mitochondrial substrates of omi indicate a new regulatory role in neurodegenerative disorders.
The mitochondrial protease OMI (also known as HtrA2) has been implicated in Parkinson's Disease (PD) and deletion or protease domain point mutations have shown profound neuropathologies in mice. A beneficial role by OMI, in preserving cell viability, is assumed to occur via the avoidance of dysfunctional protein turnover. However relatively few substrates for mitochondrial Omi are known. Here we report our identification of three novel mitochondrial substrates that impact metabolism and ATP production. Using a dual proteomic approach we have identified three interactors based upon ability to bind to OMI, and/or to persist in the proteome after OMI activity has been selectively inhibited. One candidate, the chaperone HSPA8, was common to each independent study. Two others (PDHB subunit and IDH3A subunit) did not appear to bind to OMI, however persisted in the mito-proteome when OMI was inhibited. Pyruvate dehydrogenase (PDH) and isocitrate dehydrogenase (IDH) are two key Kreb's cycle enzymes that catalyse oxidative decarboxylation control points in mitochondrial respiration. We verified both PDHB and IDH3A co-immunoprecipitate with HSPA8 and after elution, were degraded by recombinant HtrA2 in vitro. Additionally our gene expression studies, using rotenone (an inhibitor of Complex I) showed Omi expression was silenced when pdhb and idh3a were increased when a sub-lethal dose was applied. However higher dose treatment caused increased Omi expression and decreased levels of pdhb and idh3a transcripts. This implicates mitochondrial OMI in a novel mechanism relating to metabolism.
Johnson
Felicity
F
Department of Neurological Surgery, Cornell University, Weill Medical College, New York, New York, USA. fej2001@med.cornell.edu
Kaplitt
Michael G
MG
D002374:Catalase / Q000378:metabolism; D002460:Cell Line; D015971:Gene Expression Regulation, Enzymologic; D000074923:High-Temperature Requirement A Serine Peptidase 2; D006801:Humans; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D018832:Molecular Chaperones; D019636:Neurodegenerative Diseases / Q000378:metabolism*; D009474:Neurons / Q000473:pathology; D010100:Oxygen / Q000737:chemistry; D017354:Point Mutation; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D012402:Rotenone / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000378:metabolism*; D013997:Time Factors
D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; D011768:Pyruvate Dehydrogenase Complex; D012402:Rotenone; D007521:Isocitrate Dehydrogenase; D002374:Catalase; D012697:Serine Endopeptidases; C000615914:HTRA2 protein, human; D000074923:High-Temperature Requirement A Serine Peptidase 2; C000615915:Htra2 protein, mouse; D010100:Oxygen