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Site-specific photocross-linking reveals that Sec61p and TRAM contact different regions of a membrane-inserted signal sequence.

Open AccessPublished:December 15, 1993DOI:https://doi.org/10.1016/S0021-9258(19)74376-2
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      A chemically charged amber suppressor tRNA was used to introduce the photoactivatable amino acid (Tmd)Phe at a selected position within the signal sequence of the secretory protein preprolactin. This allowed the interactions of the NH2-terminal, the central, and the COOH-terminal regions of the signal sequence to be investigated during insertion into the membrane of the endoplasmic reticulum (ER). We found that different regions of the nascent chains were photocross-linked to different ER proteins. The TRAM protein (translocating chain-associating membrane protein) contacts the NH2-terminal region of the signal sequence while the mammalian Sec61p contacts the hydrophobic core of the signal sequence and regions COOH-terminal of this. These results suggest that the ER translocation complex is composed of heterologous protein subunits which contact distinct regions of nascent polypeptides during their membrane insertion.

      REFERENCES

        • Bain J.D.
        • Diala E.S.
        • Glabe C.G.
        • Dix T.A.
        • Chamberlin A.R.
        J. Am. Chem. Soc. 1989; 111: 8013-8014
        • Bain J.D.
        • Diala E.S.
        • Glabe C.G.
        • Wacker D.A.
        • Lyttle M.H.
        • Dix T.A.
        • Chamberlin A.R.
        Biochemistry. 1991; 30: 5411-5421
        • Bain J.D.
        • Wacker D.A.
        • Kuo E.E.
        • Lyttle M.H.
        • Chamberlin A.R.
        J. Org. Chem. 1991; 56: 4615-4625
        • Bain J.D.
        • Switzer C.
        • Chamberlin A.R.
        • Benner S.A.
        Nature. 1992; 356: 537-539
        • Baldini G.
        • Martoglio B.
        • Schachenmann A.
        • Zuglianin C.
        • Brunner J.
        Biochemistry. 1988; 27: 7951-7959
        • Bruce A.G.
        • Uhlenbeck O.C.
        Biochemistry. 1982; 21: 855-861
        • Brunner J.
        • Senn H.
        • Richards F.M.
        J. Biol. Chem. 1980; 255: 3313-3318
        • Chung H.-H.
        • Benson D.R.
        • Schultz P.G.
        Science. 1993; 259: 806-809
        • Deshaies R.J.
        • Schekman R.
        J. Cell Biol. 1987; 105: 633-645
        • Deshaies R.J.
        • Schekman R.
        J. Cell Biol. 1989; 109: 2653-2664
        • EUman J.A.
        • Mendel D.
        • Cahill S.A.
        • Noren C.J.
        • Schultz P.G.
        Methods Enzymol. 1991; 202: 301-336
        • Ellman J.A.
        • Volkmann B.F.
        • Mendel D.
        • Schultz P.G.
        • Wemmer D.E.
        J. Am. Chem. Soc. 1992; 114: 7959-7961
        • Ellman J.A.
        • Mendel D.
        • Schultz P.G.
        Science. 1992; 255: 197-200
        • Fujiki Y.
        • Hubbard A.L.
        • Fowler S.
        • Lazarow P.B.
        J. Cell Biol. 1982; 93: 97-102
        • Görlich D.
        • Hartmann E.
        • Prehn S.
        • Rapoport T.A.
        Nature. 1992; 357: 47-52
        • Görlich D.
        • Prehn S.
        • Hartmann E.
        • Kalies K.-U.
        • Rapoport T.A.
        Cell. 1992; 71: 489-503
        • Green N.
        • Fang H.
        • Walter P.
        J. Cell Biol. 1992; 116: 597-604
        • Haeuptle M.-T.
        • Frank R.
        • Dobberstein B.
        Nucleic Acids Res. 1986; 14: 1427-1448
        • Heckler T.G.
        • Chang L.-H.
        • Zama Y.
        • Naka T.
        • Chorghade M.S.
        • Hecht S.M.
        Biochemistry. 1984; 23: 1468-1473
        • High S.
        Bioessays. 1992; 14: 535-540
        • High S.
        • Dobberstein B.
        Neupert W. Lill R. Membrane Biogenesis and Protein Targeting-New Comprehensive Biochemistry. 22. Elsevier Science Publishers B. V., Amsterdam1992: 105-118
        • High S.
        • Görlich D.
        • Wiedmann M.
        • Rapoport T.A.
        • Dobberstein B.
        J. Cell Biol. 1991; 113: 35-44
        • High S.
        • Andersen S.S.L.
        • Görlich D.
        • Hartmann E.
        • Prehn S.
        • Rapoport T.A.
        • Dobberstein B.
        J. Cell Biol. 1993; 121: 743-750
        • Kellaris K.V.
        • Bowen S.
        • Gilmore R.
        J. Cell Biol. 1991; 114: 21-33
        • Krieg U.C.
        • Walter P.
        • Johnson A.E.
        Proc. Natl. Acad. Sci. U. S. A. 1986; 83: 8604-8608
        • Krieg U.C.
        • Johnson A.E.
        • Walter P.
        J. Cell Biol. 1989; 109: 2033-2043
        • Kurzchalia T.V.
        • Wiedmann M.
        • Girshovich A.S.
        • Bochkareva E.S.
        • Bielka H.
        • Rapoport T.A.
        Nature. 1986; 320: 634-636
        • Lipp J.
        • Dobberstein B.
        Cell. 1986; 46: 1103-1112
        • Mendel D.
        • Ellman J.A.
        • Chang Z.
        • Veenstra D.L.
        • Kollman P.A.
        • Schultz P.G.
        Science. 1992; 256: 1798-1802
        • Müsch A.
        • Wiedmann M.
        • Rapoport T.A.
        Cell. 1992; 69: 343-352
        • Nassal M.
        J. Am. Chem. Soc. 1984; 106: 7540-7545
        • Noren C.J.
        • Anthony-Cahill S.J.
        • Griffith M.C.
        • Schultz P.G.
        Science. 1989; 244: 182-188
        • Noren C.J.
        • Anthony-Cahill S.J.
        • Suich D.J.
        • Noren K.A.
        • Griffith M.C.
        • Schultz P.G.
        Nucleic Acids Res. 1989; 18: 83-88
        • Payne R.C.
        • Nichols B.P.
        • Hecht S.M.
        Biochemistry. 1987; 26: 3197-3205
        • Pezzuto J.M.
        • Hecht S.
        J. Biol. Chem. 1980; 255: 865-869
        • Rapoport T.A.
        Trends Biochem. Sci. 1990; 15: 355-358
        • Rapoport T.A.
        Science. 1992; 258: 931-936
        • Robertson S.A.
        • Ellmean J.A.
        • Schultz P.G.
        J. Am. Chem. Soc. 1991; 113: 2722-2729
        • Roesser J.R.
        • Xu C.
        • Payne R.C.
        • Surratt C.K.
        • Hecht S.M.
        Biochemistry. 1989; 28: 5185-5195
        • Rothblatt J.A.
        • Deshaies R.J.
        • Sanders S.L.
        • Daum G.
        • Schekman R.
        J. Cell Biol. 1989; 109: 2641-2652
        • Sadler I.
        • Chiang A.
        • Kurihara T.
        • Rothblatt J.
        • Way J.
        • Silver P.
        J. Cell Biol. 1989; 109: 2665-2675
        • Sanders S.L.
        • Whitfield K.M.
        • Vogel J.P.
        • Rose M.D.
        • Schekman R.W.
        Cell. 1992; 69: 353-365
        • Siegel V.
        • Walter P.
        Cell. 1988; 52: 39-49
        • Simon S.M.
        • Blobel G.
        Cell. 1991; 65: 371-380
        • Stirling C.J.
        • Rothblatt J.
        • Hosobuchi H.
        • Deshaies R.
        • Schekman R.
        Mol. Biol. Cell. 1992; 3: 129-142
        • Thrift R.N.
        • Andrews D.W.
        • Walter P.
        • Johnson A.E.
        J. Cell Biol. 1991; 112: 809-821
        • von Heijne G.
        Biochim. Biophys. Acta. 1988; 947: 307-333
        • Wiedmann M.
        • Kurzchalia T.V.
        • Hartmann E.
        • Rapoport T.A.
        Nature. 1987; 328: 830-833
        • Wiedmann M.
        • Görlich D.
        • Hartmann E.
        • Kurzchalia T.V.
        • Rapoport T.A.
        FEBS Lett. 1989; 257: 263-268