title: The influence of depletion of voltage dependent anion selective channel on protein import into the yeast Saccharomyces cerevisiae mitochondria.
abstract: The supply of substrates to the respiratory chain as well as of other metabolites (e.g. ATP) into inner compartments of mitochondria is crucial to preprotein import into these organelles. Transport of the compounds across the outer mitochondrial membrane is enabled by mitochondrial porin, also known as the voltage-dependent anion-selective channel (VDAC). Our previous studies led to the conclusion that the transport of metabolites through the outer membrane of the yeast Saccharomyces cerevisiae mitochondria missing VDAC (now termed YVDAC1) is considerably restricted. Therefore we expected that depletion of YVDAC1 should also hamper protein import into the mutant mitochondria. We report here that YVDAC1-depleted mitochondria are able to import a fusion protein termed pSu9-DHFR in the amount comparable to that of wild type mitochondria, although over a considerably longer time. The rate of import of the fusion protein into YVDAC1-depleted mitochondria is dis- tinctly lower than into wild type mitochondria probably due to restricted ATP access to the intermembrane space and is additionally influenced by the way the supporting respiratory substrates are transported through the outer membrane. In the presence of ethanol, diffusing freely through lipid membranes, YVDAC1-depleted mitochondria are able to import the fusion protein at a higher rate than in the presence of external NADH which is, like ATP, transported through the outer membrane by facilitated diffusion. It has been shown that transport of external NADH across the outer membrane of YVDAC1-depleted mitochondria is supported by the protein import machinery, i.e. the TOM complex (Kmita & Budzińska, 2000, Biochim. Biophys. Acta 1509, 86-94.). Since the TOM complex might also contribute to the permeability of the membrane to ATP, it seems possible that external NADH and ATP as well as the imported preprotein could compete with one another for the passage through the outer membrane in YVDAC1-depleted mitochondria.
authors.affiliation: Institute of Molecular Biology and Biotechnology, Department of Bioenergetics, Poznań University, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Szczechowicz
authors.forename: L
authors.initials: L
authors.lastname: Hryniewiecka
authors.forename: H
authors.initials: H
authors.lastname: Kmita
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001693:Biological Transport, Active; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D018272:Porins / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D050994:Voltage-Dependent Anion Channels
chemical_list: D018272:Porins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D050994:Voltage-Dependent Anion Channels; D000255:Adenosine Triphosphate