title: Inactivation of the Neurospora crassa mitochondrial outer membrane protein TOM70 by repeat-induced point mutation (RIP) causes defects in mitochondrial protein import and morphology.
abstract: Mitochondrial biogenesis requires the efficient import of hundreds of different cytosolically translated preproteins into existing organelles. Recognition and translocation of preproteins at the mitochondrial outer membrane is achieved by the TOM complex (translocase of the outer mitochondrial membrane). The largest component of this complex is TOM70, an integral outer membrane protein with a large cytosolic domain thought to serve as a receptor for a specific group of preproteins. To investigate the functional role of TOM70 in Neurospora crassa the tom70 gene was inactivated using the natural phenomenon of repeat-induced point mutation (RIP). Mutant strains were identified that harbored RIPed tom70 alleles and contained no immunologically detectable TOM70. Strains that lack TOM70 grow more slowly than wild-type strains, conidiate poorly, and contain enlarged mitochondria. In vitro preprotein import studies using TOM70-deficient mitochondria revealed a defect in the uptake of the ADP/ATP carrier. Other preproteins tested were imported at wild-type rates with the exception of the precursor of the mitochondrial-processing peptidase (MPP) which was imported more efficiently by TOM70-deficient mitochondria. These data support the view that TOM70 plays a role as a specific receptor for carrier proteins in mitochondrial-preprotein import. The presence of tetratricopeptide repeats (TPRs) in the TOM70 sequence and the enlarged shape of mitochondria lacking TOM70 raise the possibility that the protein also plays a role in the maintenance of mitochondrial morphology.
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
authors.forename: L I
authors.initials: LI
authors.lastname: Grad
authors.forename: A T
authors.initials: AT
authors.lastname: Descheneau
authors.forename: W
authors.initials: W
authors.lastname: Neupert
authors.forename: R
authors.initials: R
authors.lastname: Lill
authors.forename: F E
authors.initials: FE
authors.lastname: Nargang
mesh_terms: D002455:Cell Division; D019286:Endopeptidase K / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology*; D008854:Microscopy, Electron; D008928:Mitochondria / Q000737:chemistry* / Q000648:ultrastructure; D009492:Neurospora crassa / Q000235:genetics* / Q000502:physiology*; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D017354:Point Mutation; D014357:Trypsin / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D008565:Membrane Proteins; D014357:Trypsin; D019286:Endopeptidase K