The influence of depletion of voltage dependent anion selective channel on protein import into the yeast Saccharomyces cerevisiae mitochondria

• Autorzy: Szczechowicz A. , Hryniewiecka L. , Kmita H.
• Języki publikacji: EN

Abstrakty

EN

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 an- ion-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 YVD AC 1) is considerably re­stricted. Therefore we expected that depletion of YVDAC1 should also hamper pro­tein import into the mutant mitochondria. We report here that YVD AC 1-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 YVD AC 1-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 mitochon- dria 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 machin- ery, i.e. the TOM complex (Kmita & Budziñska, 2000, Biochim. Biophys. Acta 1509, 86.94.). Since theTOMcomplex might also contribute to the permeability of themem-brane 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 mem- brane in YVDAC1-depleted mitochondria.

Słowa kluczowe

EN

metabolite; yeast; anion selective channel; metabolite transport; mitochondrion; respiratory chain; protein import; mitochondrial membrane; protein; depletion; Saccharomyces cerevisiae

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2001
• Tom: 48
• Numer: 3
• Opis fizyczny: p.719-728,fig.

Twórcy

autor

Szczechowicz A.

• Poznan University, A.Fredry 10, 61-701 Poznan, Poland

autor

Hryniewiecka L.

autor

Kmita H.

Bibliografia

• 1.Lill, R. & Neupert, W. (1996) Mechanisms of protein import across the mitochondrial outer membrane. Trends Cell Biol. 6, 56-61.
• 2.Pfanner, N. & Meijer, M. (1997) The Tom and Tim machine. Curr. Biol. 7, R100-R103.
• 3.Pfanner, N. (1998) Crossing the aqueous intermembrane space. Curr. Biol. 8, R262-R265.
• 4.Schein, S.J., Colombini, M. & Finkelstein, A. (1976) Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel. J. Membr. Biol. 30, 99-120.
• 5.Colombini, M. (1979) A candidate for the permeability pathway of the outer mitochondrial membrane. Nature 279, 643-645.
• 6.Benz, R. (1994) Permeation of hydrophilic solutes through mitochondrial outer membranes: Review on mitochondrial porins. Biochim. Biophys. Acta 1197, 167-196.
• 7.Colombini, M., Blachly-Dyson, E. & Forte, M. (1996) VDAC, a channel in the outer mitochondrial membrane; in Ion Channels (Narahashi, T., ed.) vol. 4, pp. 169-201, Plenum Press, New York.
• 8.Guo, X.J. & Lauquin, G.J.-M. (1986) Mitochondrial porin-deficient mutant of S. cerevisiae: In vitro construction and preliminary properties. EBEC Reports 4, 292.
• 9.Dihanich, M., Suda, K. & Schatz, G. (1987) A yeast mutant lacking mitochondrial porin is respiratory-deficient, but can recover respiration with simultaneous accumulation of an 86kD extramitochondrial protein. EMBO J. 6, 723-728.
• 10.Blachly-Dyson, E., Song, J., Wolfgand, W.J., Colombini, M. & Forte, M. (1997) Multicopy suppressors of phenotype resulting from the absence of yeast VDAC encode a VDAC-like protein. Mol. Cell. Biol. 17, 5727-5738.
• 11.Dihanich, M., Schmid, A., Oppliger, W. & Benz, R. (1989) Identification of a new pore in the mitochondrial outer membrane of a porin- deficient yeast mutant. Eur. J. Biochem. 181, 703-708.
• 12.Benz, R., Schmid, A. & Dihanich, M. (1989) Pores from mitochondrial outer membrane of yeast and a porin-deficient yeast mutant: A comparison. J. Bioener. Biomembr. 21, 439- 450.
• 13.Michejda, J., Kmita, H., Stobienia, O., Budzinska, M. & Lauquin, G.J.-M. (1994) Restriction of metabolite permeation through the outer mitochondrial membrane of porin-deficient yeast mutant; in Molecular Biology of Mitochondrial Transport System (Forte, M. & Colombini, M., eds.) pp. 341-356, Springer-Verlag, Heidelberg.
• 14.Lee, A.C., Xu, X., Blachly-Dyson, E., Forte, M. & Colombini, M. (1998) The role of yeast VDAC genes on the permeability of the mitochondrial outer membrane. J. Membr. Biol. 161, 173-181.
• 15.Kmita, H., Stobienia, O. & Michejda, J. (1999) The access of metabolites into yeast mitochondria in the presence and absence of the voltage dependent anion selective channel (YVDAC1). Acta Biochim. Polon. 46, 991-1000.
• 16.Thieffry, M., Chich, J.M., Goldschmidt, D. & Henry, J.P. (1988) Incorporation in lipid bilayers of a large conductance cationic channel from mitochondrial membranes. EMBO J. 7, 1449-1454.
• 17.Fevre, F., Chich, J.F., Lauquin, G.J.-M., Henry, J.P. & Thieffry, M. (1990) Comparison of mitochondrial cationic channels in wild- type and porin-deficient mutant yeast. FEBS Lett. 262, 201-204.
• 18.Kuenkele, K.-P., Heins, S., Dembowski, M., Nargang, F.E., Benz, R., Thieffry, M., Walz, J., Lill, R., Nussberger, S. & Neupert, W. (1998) The preprotein translocation channel of the outer membrane of mitochondria. Cell 93, 1009-1019.
• 19.Kuenkele, K.-P., Juin, P., Pompa, C., Nargang, F.E., Henry, J.-P., Neupert, W., Lill, R. & Thieffry, M. (1998) The isolated complex of the translocase of the outer membrane of mitochondria. J. Biol. Chem. 273, 31032-31039.
• 20.Hill, K., Model, K., Ryan, M.T., Dietmeier, K., Martin, F., Wagner, R. & Pfanner, N. (1998) Tom40 forms the hydrophilic channel of the mitochondrial import pore for preproteins. Nature 395, 516-521.
• 21.Vallette, F.M., Juin, P., Pelleschi, M. & Henry, J.-P. (1994) Basic peptides can be imported into yeast mitochondria by two distinct pathways. J. Biol. Chem. 269, 13367-13374.
• 22.Juin, P., Thieffry, M., Henry, J.-P. & Vallette, F.M. (1997) Relationship between the peptide-sensitive channel and the mitochondrial outer membrane protein translocation machinery. J. Biol. Chem. 272, 6044-6050.
• 23.Kmita, H., Budzinska, M. & Michejda, J. (1999) NADH and proteins may use the same pathway to cross the outer membrane in VDAC-depleted mitochondria. Biol. Bull. Poznan 36, 5-15.
• 24.Kmita, H. & Budzinska, M. (2000) Involvement of the TOM complex in external NADH transport into yeast mitochondria depleted of porin 1. Biochim. Biophys. Acta 1509, 86-94.
• 25.Michejda, J., Guo, X.J. & Lauquin, G.J.-M. (1989) Bioenergetic consequences of the lack of mitochondrial porin: Identification of a putative new pore; in Anion Carriers of Mitochondrial Membranes (Azzi, A., Nalecz, K.A., Nalecz, M.J. & Wojtczak, L., eds.) pp. 225-235, Springer-Verlag, Berlin, Heidelberg.
• 26.Daum, G., Bohni, P.C. & Schatz, G. (1982) Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J. Biol. Chem. 257, 13028-13033.
• 27.Douce, R., Bourguignon, J., Brouquisse, R. & Neuberger, M. (1984) Isolation of plant mitochondria. General principles and criteria of integrity. Methods Enzymol. 148, 403-415.
• 28.Pfanner, N., Mueller, H.K., Harmey, M.A. & Neupert, W. (1987) Mitochondrial protein import: Involvement of the mature part of a cleavable precursor protein in the binding to receptor sites. EMBO J. 6, 3449-3454.
• 29.Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.
• 30.Kamo, N., Maratsugu, M., Hongoh, R. & Kobatake, Y.J. (1979) Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential and phosphorylation potential in steady state. J. Membr. Biol. 49, 105-121.
• 31.Chance, B. & Williams, G.R. (1956) The respiratory chain and oxidative phosphorylation. Adv. Enzymol. 17, 65-87.
• 32.de Vries, S. & Marres, C.A.M. (1991) The mitochondrial respiratory chain of yeast. Structure and biosynthesis and the role in cellular metabolism. Biochim. Biophys. Acta 895, 205-239.
• 33.Michejda, J., Guo, X.J. & Lauquin, G.J.-M. (1990) The respiration of cells and mitochondria of porin deficient yeast mutants is coupled. Biochem. Biophys. Res. Commun. 171, 354-361.