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Soluble and Membrane-Associated Factors Required for Protein Translocation into the Yeast Endoplasmic Reticulum

  • Conference paper
Dynamics and Biogenesis of Membranes

Part of the book series: NATO ASI Series ((ASIH,volume 40))

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Summary

A genetic approach aimed at defining molecules that participate in protein translocation across the membrane of the endoplasmic reticulum (ER) has led to the identification of three new genes. Mutant cells that fail to translocate secretory proteins into the lumen of the ER have been selected by a procedure that demands retention of a signal peptide-containing hybrid protein in the cytosol. Temperature sensitive mutations were obtained in which precursors of secreted and vacuolar proteins accumulate in forms that are devoid of any posttranslational modifications normally coupled to translocation into the ER. Pre-proteins that accumulate at the mutant block sediment with membranes, but are exposed to the cytosol as judged by sensitivity to exogenous protease. The precursor of α-factor is arrested in a form that can complete translocation and secretion during an extended period of chase, confirming in vitro results that show protein synthesis and membrane assembly are not obligately coupled.

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References

  • Akiyama Y and Ito K (1987) Topology analysis of the SecY protein, an integral membrane protein involved in protein export in Escherichia coll. EMBO J. 6: 3465–3470

    PubMed  CAS  Google Scholar 

  • Basson ME, Thorsness M, Finer-Moore J, Stroud RM and Rine J (1988) Structural and functional conservation between yeast and human 3-hydroxy-3-methylglutaryl coenzyme A reductases. Mol. Cell. Biol. 8: 3797–3808

    CAS  Google Scholar 

  • Bernstein M, Kepes F and Schekman R (1989) SEC59 encodes a membrane protein required for core glycosylation in Saccharomyces cerevisiae. Mol. Cell. Biol. 9: 1191–1199

    PubMed  CAS  Google Scholar 

  • Chappell TG, Welch WJ, Schlossman DM, Palter KB, Schlesinger MJ and Rothman JE (1986) Uncoating ATPase is a member of the 70 kilodalton family of stress proteins. Cell 45: 3–13

    Article  PubMed  CAS  Google Scholar 

  • Chen W-J and Douglas MG (1987a) Phosphodiester bond cleavage outside mitochondria is required for the completion of protein import into the mitochondrial matrix. Cell 49: 651–658

    Article  PubMed  CAS  Google Scholar 

  • Chen W-J and Douglas MG (1987b) The role of protein structure in the mitochondrial import pathway. J. Biol. Chem. 262: 15598–15604

    PubMed  CAS  Google Scholar 

  • Chirico WJ, Waters MG and Blobel G (1988) 70 K heat shock related proteins stimulate protein translocation into microsomes. Nature 332: 805–810

    Article  PubMed  CAS  Google Scholar 

  • Craig EA, Kramer J, and Kosic-Smithers J (1987) SSC1, a member of the 70-kDa heat shock protein multigene family of Saccharomyces cerevisiae, is essential for growth. Proc. Natl. Acad. Sci. USA 84:4156–4160

    Article  PubMed  CAS  Google Scholar 

  • Crooke E, Guthrie B, Lecker S, Lill R and Wickner W (1988) proOmpA is stabilized for membrane translocation by either purified E. coli trigger factor or canine SRP. Cell 54: 1003–1011

    Article  PubMed  CAS  Google Scholar 

  • Deshaies RJ and Schekman R (1987) A yeast mutant defective at an early stage in import of secretory protein precursors into the endoplasmic reticulum. J. Cell Biol. 105: 633–645

    Article  PubMed  CAS  Google Scholar 

  • Deshaies RJ, Koch BD, Werner-Washburne M, Craig EA, and Schekman R (1988) 70 kD stress protein homologues facilitate translocation of secretory and mitochondrial precursor polypeptides. Nature 332: 800–805

    Article  PubMed  CAS  Google Scholar 

  • Eilers M and Schatz G (1986) Binding of a specific ligand inhibits import of a purified precursor protein into mitochondria. Nature 332: 228–232

    Article  Google Scholar 

  • Eilers M, Ipplinger W and Schatz G (1987) Both ATP and an energized inner membrane are required to import a purified precursor protein into mitochondria. EMBO J 6: 1073–1077

    PubMed  CAS  Google Scholar 

  • Emr SD, Schauer I, Hansen W, Esmon P and Schekman R (1984) Invertase-(β-galactosidase hybrid proteins fail to be transported from the endoplasmic reticulum in Saccharomyces cerevisiae. Mol. Cell. Biol. 4: 2347–2355

    PubMed  CAS  Google Scholar 

  • Feldman RI, Bernstein M and Schekman R (1987) Product of SEC53 is required for folding and glycosylation of secretory proteins in the lumen of the yeast endoplasmic reticulum. J. Biol. Chem. 262: 9332–9339

    PubMed  CAS  Google Scholar 

  • Ferro-Novick S, Novick P, Field C and Schekman R (1984a) Yeast secretory mutants that block the formation of active cell surface enzymes. J Cell Biol. 98: 35–43

    Article  PubMed  CAS  Google Scholar 

  • Hansen W, Garcia PD and Walter P (1986) in vitro translocation across the yeast endoplasmic reticulum: ATP-dependent post-translational translocation of the prepro-α-factor. Cell 45: 397–406

    Article  PubMed  CAS  Google Scholar 

  • Keesey JK, Bigelis R, and Fink GR (1979) The product of the his4 gene cluster in Saccharomyces cerevisiae. J. Biol. Chem. 254: 7427–7433

    PubMed  CAS  Google Scholar 

  • Lewis MJ and Pelham HRB (1985) Involvement of ATP in the nuclear and nucleolar functions of the 70 kd heat shock protein. EMBO J. 4: 3137–3143

    PubMed  CAS  Google Scholar 

  • Munro S and Pelham HRB (1986) An hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell 46: 291–300

    Article  PubMed  CAS  Google Scholar 

  • Murakami H, Pain D and Blobel G (1988) 70-kD heat shock-related protein is one of at least two distinct cytosolic factors stimulating protein import into mitochondria. J. Cell Biol. 107: 2051–2057

    Article  PubMed  CAS  Google Scholar 

  • Novick P and Schekman R (1979) Secretion and cell-surface growth are blocked in a temperature-sensitive mutant of Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 76: 1858–1862

    Article  PubMed  CAS  Google Scholar 

  • Novick P, Field C and Schekman R (1980) Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell 21: 202–215

    Article  Google Scholar 

  • Ohta S and Schatz G (1984) A purified precursor polypeptide requires a cytosolic protein fraction for import into mitochondria. EMBO J 3: 651–657

    PubMed  CAS  Google Scholar 

  • Parker R and Guthrie C (1985) A point mutation in the conserved hexanucleotide at a yeast 5’ splice junction uncouples recognition, cleavage, and ligation. Cell 41: 107–118

    Article  PubMed  CAS  Google Scholar 

  • Pelham HRB (1986) Speculations on the functions of the major heat shock and glucose- regulated proteins. Cell 46: 959–961

    Article  PubMed  CAS  Google Scholar 

  • Pfanner N, Tropschug M and Neupert W (1987) Mitochondrial protein import: nucleoside triphosphates are involved in conferring import-competence to precursors. Cell 49: 815–823

    Article  PubMed  CAS  Google Scholar 

  • Rothblatt JA and Meyer DI (1986a) Secretion in yeast: reconstitution of the translocation and glycosylation of a-factor and invertase in a homologous cell-free system. Cell 44: 619–628

    Article  PubMed  CAS  Google Scholar 

  • Rothblatt J and Meyer DI (1986b) Secretion in yeast: translocation and glycosylation of prepro- α-factor in vitro can occur via an ATP-dependent post-translational mechanism. EMBO J. 5: 1031–1036

    PubMed  CAS  Google Scholar 

  • Rothman JE and Kornberg RD (1986) An unfoloding story of protein translocation. Nature 322: 209–210

    Article  PubMed  CAS  Google Scholar 

  • Sanz P and Meyer DI (1988) Signal recognition particle (SRP) stabilizes the translocation- competent conformation of pre-secretory proteins. EMBO J 7: 3553–3557

    PubMed  CAS  Google Scholar 

  • Schlossman DM, Schmid SL, Braell WA and Rothman JE (1984) An enzyme that removes clathrin coats: purification of an uncoating ATPase. J Cell Biol. 99: 723–733

    Article  PubMed  CAS  Google Scholar 

  • Sharma S, Rodgers L, Brandsma J, Gething M-J and Sambrook J (1985) SV40 T antigen and the exocytotic pathway. EMBO J. 4: 1479–1489

    PubMed  CAS  Google Scholar 

  • Simon K, Perara E and Lingappa, VR (1987) Translocation of globin fusion proteins across the endoplasmic reticulum membrane in Xenopus laevis oocytes. J. Cell Biol. 104: 1165–1172

    Article  PubMed  CAS  Google Scholar 

  • Ungewickell E (1985) The 70-kd mammalian heat shock proteins are structurally and functionally related to the uncoating protein that releases clathrin triskelia from coated vesicles. EMBO J 4: 3385–3391

    PubMed  CAS  Google Scholar 

  • Waters MG and Blobel G (1986) Secretory protein translocation in a yeast cell-free system can occur post-translationally and requires ATP hydrolysis. J. Cell Biol. 102: 1543–1550

    Article  PubMed  CAS  Google Scholar 

  • Waters MG, Chirico WJ and Blobel G (1986) Protein translocation across the yeast microsomal membrane is stimulated by a soluble factor. J. Cell Biol. 103:2629–2636

    Article  PubMed  CAS  Google Scholar 

  • Werner-Washburne M, Stone DE and Craig EA (1987) Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae. Mol. Cell Biol. 7: 2568–2577

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, University of California, 94720, Berkeley, CA, USA

    R. J. Deshaies, A. Eun, B. D. Koch, J. A. Rothblatt, S. Sanders, C. Stirling & R. Schekman

Authors
  1. R. J. Deshaies
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  2. A. Eun
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  3. B. D. Koch
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  4. J. A. Rothblatt
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  5. S. Sanders
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  6. C. Stirling
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  7. R. Schekman
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Editor information

Editors and Affiliations

  1. Centre for Biomembranes and Lipid Enzymology, Padualaan 8, 3584 CH, Utrecht, The Netherlands

    J. A. F. Op den Kamp

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© 1990 Springer-Verlag Berlin Heidelberg

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Deshaies, R.J. et al. (1990). Soluble and Membrane-Associated Factors Required for Protein Translocation into the Yeast Endoplasmic Reticulum. In: Op den Kamp, J.A.F. (eds) Dynamics and Biogenesis of Membranes. NATO ASI Series, vol 40. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-74194-4_24

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  • DOI: https://doi.org/10.1007/978-3-642-74194-4_24

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-74196-8

  • Online ISBN: 978-3-642-74194-4

  • eBook Packages: Springer Book Archive

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