Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70

doi:10.1083/jcb.120.1.95

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Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70

JL Brodsky, S Hamamoto, D Feldheim and R Schekman
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley 94720.

We reconstituted prepro-alpha-factor translocation and signal peptide processing using a yeast microsomal detergent soluble fraction formed into vesicles with soybean phospholipids. Reconstituted translocation required ATP, and was deficient when sec63 and kar2 (BiP) mutant cells were used as a source of membranes. Normal translocation was observed with vesicles reconstituted from a mixture of pure wild-type yeast BiP and a soluble fraction of kar2 mutant membranes. Two other heat-shock cognate (hsc) 70 homologs, yeast cytosolic hsc70 (Ssalp) and E. coli dnaK protein did not replace BiP. Conversely, BiP was not active under conditions where translocation into native ER vesicles required cytosolic hsc70. We conclude that cytosolic hsc70 and BiP serve noninterchangeable roles in polypeptide translocation, possibly because distinct, asymmetrically oriented membrane proteins are required to recruit each protein to opposing surfaces of the ER membrane.
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Brown, C. R., McCann, J. A., Chiang, H.-L. (2000). The Heat Shock Protein Ssa2p Is Required for Import of Fructose-1,6-Bisphosphatase into Vid Vesicles. JCB 150: 65-76 [Abstract] [Full Text]
Oh, H. J., Easton, D., Murawski, M., Kaneko, Y., Subjeck, J. R. (1999). The Chaperoning Activity of hsp110. IDENTIFICATION OF FUNCTIONAL DOMAINS BY USE OF TARGETED DELETIONS. J. Biol. Chem. 274: 15712-15718 [Abstract] [Full Text]
Brodsky, J. L., Werner, E. D., Dubas, M. E., Goeckeler, J. L., Kruse, K. B., McCracken, A. A. (1999). The Requirement for Molecular Chaperones during Endoplasmic Reticulum-associated Protein Degradation Demonstrates That Protein Export and Import Are Mechanistically Distinct. J. Biol. Chem. 274: 3453-3460 [Abstract] [Full Text]
Pilon, M., Römisch, K., Quach, D., Schekman, R. (1998). Sec61p Serves Multiple Roles in Secretory Precursor Binding and Translocation into the Endoplasmic Reticulum Membrane. Mol. Biol. Cell 9: 3455-3473 [Abstract] [Full Text]
McClellan, A. J., Endres, J. B., Vogel, J. P., Palazzi, D., Rose, M. D., Brodsky, J. L. (1998). Specific Molecular Chaperone Interactions and an ATP-dependent Conformational Change Are Required during Posttranslational Protein Translocation into the Yeast ER. Mol. Biol. Cell 9: 3533-3545 [Abstract] [Full Text]
Lu, Z., Cyr, D. M. (1998). Protein Folding Activity of Hsp70 Is Modified Differentially by the Hsp40 Co-chaperones Sis1 and Ydj1. J. Biol. Chem. 273: 27824-27830 [Abstract] [Full Text]
Chevalier, M., King, L., Wang, C., Gething, M.-J., Elguindi, E., Blond, S. Y. (1998). Substrate Binding Induces Depolymerization of the C-terminal Peptide Binding Domain of Murine GRP78/BiP. J. Biol. Chem. 273: 26827-26835 [Abstract] [Full Text]
Paunola, E., Suntio, T., Jämsä, E., Makarow, M. (1998). Folding of Active beta -Lactamase in the Yeast Cytoplasm before Translocation into the Endoplasmic Reticulum. Mol. Biol. Cell 9: 817-827 [Abstract] [Full Text]
Lu, Z., Cyr, D. M. (1998). The Conserved Carboxyl Terminus and Zinc Finger-like Domain of the Co-chaperone Ydj1 Assist Hsp70 in Protein Folding. J. Biol. Chem. 273: 5970-5978 [Abstract] [Full Text]
Shieh, H.-L., Chiang, H.-L. (1998). In Vitro Reconstitution of Glucose-induced Targeting of Fructose-1,6-bisphosphatase into the Vacuole in Semi-intact Yeast Cells. J. Biol. Chem. 273: 3381-3387 [Abstract] [Full Text]
Coury, L. A., Mathai, J. C., Prasad, G. V.�R., Brodsky, J. L., Agre, P., Zeidel, M. L. (1998). Reconstitution of water channel function of aquaporins 1�and 2�by expression in yeast secretory vesicles. Am. J. Physiol. Renal Physiol. 274: F34-F42 [Abstract] [Full Text]
Corsi, A. K., Schekman, R. (1997). The Lumenal Domain of Sec63p Stimulates the ATPase Activity of BiP and Mediates BiP Recruitment to the Translocon in Saccharomyces cerevisiae. JCB 137: 1483-1493 [Abstract] [Full Text]
Belden, W. J., Barlowe, C. (1996). Erv25p, a Component of COPII-coated Vesicles, Forms a Complex with Emp24p That Is Required for Efficient Endoplasmic Reticulum to Golgi Transport. J. Biol. Chem. 271: 26939-26946 [Abstract] [Full Text]
King, C., Eisenberg, E., Greene, L. (1995). Polymerization of 70-kDa Heat Shock Protein by Yeast DnaJ in ATP. J. Biol. Chem. 270: 22535-22540 [Abstract] [Full Text]
Wawrzyn�w, A., Zylicz, M. (1995). Divergent Effects of ATP on the Binding of the DnaK and DnaJ Chaperones to Each Other, or to Their Various Native and Denatured Protein Substrates. J. Biol. Chem. 270: 19300-19306 [Abstract] [Full Text]
Mayer, A., Neupert, W., Lill, R. (1995). Translocation of Apocytochrome c across the Outer Membrane of Mitochondria. J. Biol. Chem. 270: 12390-12397 [Abstract] [Full Text]
Wall, D., Zylicz, M., Georgopoulos, C. (1995). The Conserved G/F Motif of the DnaJ Chaperone Is Necessary for the Activation of the Substrate Binding Properties of the DnaK Chaperone. J. Biol. Chem. 270: 2139-2144 [Abstract] [Full Text]
Simon, S.M. (1995). Protein-conducting Channels for the Translocation of Proteins into and across Membranes. Cold Spring Harb Symp Quant Biol 60: 57-69 [Abstract]
Gething, M.-J., Blond-Elguindi, S., Buchner, J., Fourie, A., Knarr, G., Modrow, S., Nanu, L., Segal, M., Sambrook, J. (1995). Binding Sites for Hsp70 Molecular Chaperones in Natural Proteins. Cold Spring Harb Symp Quant Biol 60: 417-428 [Abstract]
Wickner, W. (1994). How ATP drives proteins across membranes. Science 266: 1197-1198
Chevalier, M., Rhee, H., Elguindi, E. C., Blond, S. Y. (2000). Interaction of Murine BiP/GRP78 with the DnaJ Homologue MTJ1. J. Biol. Chem. 275: 19620-19627 [Abstract] [Full Text]