title: Biogenesis of the yeast frataxin homolog Yfh1p. Tim44-dependent transfer to mtHsp70 facilitates folding of newly imported proteins in mitochondria.
abstract: Tim44 is an essential component of the mitochondrial inner membrane protein import machinery. In this study we asked if Tim44 is of relevance in intramitochondrial protein folding. We investigated the role of Tim44 in the biogenesis of the authentic mitochondrial protein Yfh1p, the yeast homolog of mammalian frataxin, which was recently implicated in Friedreich ataxia. After inactivation of Tim44, binding of mitochondrial heat shock protein (mtHsp)70 to translocating Yfh1p and subsequent folding to the native state was nearly completely blocked. Residual amounts of imported Yfh1p showed an increased tendency to aggregate. To further characterize the functions of Tim44 in the matrix, we imported dihydrofolate reductase (DHFR) as a model protein. Depletion of Tim44 allowed import of DHFR, although folding of the newly imported DHFR was delayed. Moreover, the depletion of Tim44 caused a strongly reduced binding of mtHsp70 and Mge1 to the translocating polypeptide. Subsequent dissociation of mtHsp70 from imported DHFR was delayed, indicating that mtHsp70-substrate complexes formed independently of Tim44 differ from the complexes that form under the control of Tim44. We conclude that Tim44 not only plays a role in protein translocation but also in the pathways of mitochondrial protein folding.
authors.affiliation: Institut für Biochemie und Molekularbiologie, Universität Freiburg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Geissler
authors.forename: T
authors.initials: T
authors.lastname: Krimmer
authors.forename: B
authors.initials: B
authors.lastname: Schönfisch
authors.forename: M
authors.initials: M
authors.lastname: Meijer
authors.forename: J
authors.initials: J
authors.lastname: Rassow
mesh_terms: D001692:Biological Transport; D002352:Carrier Proteins / Q000502:physiology*; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000502:physiology*; D007425:Intracellular Membranes / Q000378:metabolism; D033862:Iron-Binding Proteins; D008565:Membrane Proteins / Q000502:physiology*; D008928:Mitochondria / Q000378:metabolism*; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis* / Q000737:chemistry; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry; D000096923:Frataxin
chemical_list: D002352:Carrier Proteins; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; D033862:Iron-Binding Proteins; D008565:Membrane Proteins; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D029701:Saccharomyces cerevisiae Proteins; D013762:Tetrahydrofolate Dehydrogenase; D000096923:Frataxin; C077016:TIM44 protein, S cerevisiae