• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Aug 15, 1997; 16(16): 4880–4886.
PMCID: PMC1170123

Co-translational protein targeting catalyzed by the Escherichia coli signal recognition particle and its receptor.

Abstract

The Ffh-4.5S ribonucleoprotein particle (RNP) and FtsY from Escherichia coli are homologous to essential components of the mammalian signal recognition particle (SRP) and SRP receptor, respectively. The ability of these E. coli components to function in a bona fide co-translational targeting pathway remains unclear. Here we demonstrate that the Ffh-4.5S RNP and FtsY can efficiently replace their mammalian counterparts in targeting nascent secretory proteins to microsomal membranes in vitro. Targeting in the heterologous system requires a hydrophobic signal sequence, utilizes GTP and, moreover, occurs co-translationally. Unlike mammalian SRP, however, the Ffh-4.5S RNP is unable to arrest translational elongation, which results in a narrow time window for the ribosome nascent chain to interact productively with the membrane-bound translocation machinery. The highly negatively charged N-terminal domain of FtsY, which is a conserved feature among prokaryotic SRP receptor homologs, is important for translocation and acts to localize the protein to the membrane. Our data illustrate the extreme functional conservation between prokaryotic and eukaryotic SRP and SRP receptors and suggest that the basic mechanism of co-translational protein targeting is conserved between bacteria and mammals.

Full Text

The Full Text of this article is available as a PDF (286K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andrews DW, Lauffer L, Walter P, Lingappa VR. Evidence for a two-step mechanism involved in assembly of functional signal recognition particle receptor. J Cell Biol. 1989 Mar;108(3):797–810. [PMC free article] [PubMed]
  • Bassford P, Beckwith J, Ito K, Kumamoto C, Mizushima S, Oliver D, Randall L, Silhavy T, Tai PC, Wickner B. The primary pathway of protein export in E. coli. Cell. 1991 May 3;65(3):367–368. [PubMed]
  • Bernstein HD, Poritz MA, Strub K, Hoben PJ, Brenner S, Walter P. Model for signal sequence recognition from amino-acid sequence of 54K subunit of signal recognition particle. Nature. 1989 Aug 10;340(6233):482–486. [PubMed]
  • Bernstein HD, Zopf D, Freymann DM, Walter P. Functional substitution of the signal recognition particle 54-kDa subunit by its Escherichia coli homolog. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5229–5233. [PMC free article] [PubMed]
  • Brown S. Time of action of 4.5 S RNA in Escherichia coli translation. J Mol Biol. 1989 Sep 5;209(1):79–90. [PubMed]
  • Bukau B, Hesterkamp T, Luirink J. Growing up in a dangerous environment: a network of multiple targeting and folding pathways for nascent polypeptides in the cytosol. Trends Cell Biol. 1996 Dec;6(12):480–486. [PubMed]
  • Bult CJ, White O, Olsen GJ, Zhou L, Fleischmann RD, Sutton GG, Blake JA, FitzGerald LM, Clayton RA, Gocayne JD, et al. Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii. Science. 1996 Aug 23;273(5278):1058–1073. [PubMed]
  • Connolly T, Gilmore R. Formation of a functional ribosome-membrane junction during translocation requires the participation of a GTP-binding protein. J Cell Biol. 1986 Dec;103(6 Pt 1):2253–2261. [PMC free article] [PubMed]
  • de Gier JW, Mansournia P, Valent QA, Phillips GJ, Luirink J, von Heijne G. Assembly of a cytoplasmic membrane protein in Escherichia coli is dependent on the signal recognition particle. FEBS Lett. 1996 Dec 16;399(3):307–309. [PubMed]
  • Fisher DK, Higgins TJ. A sensitive, high-volume, colorimetric assay for protein phosphatases. Pharm Res. 1994 May;11(5):759–763. [PubMed]
  • Freymann DM, Keenan RJ, Stroud RM, Walter P. Structure of the conserved GTPase domain of the signal recognition particle. Nature. 1997 Jan 23;385(6614):361–364. [PubMed]
  • Gill DR, Salmond GP. The identification of the Escherichia coli ftsY gene product: an unusual protein. Mol Microbiol. 1990 Apr;4(4):575–583. [PubMed]
  • Gilmore R, Blobel G, Walter P. Protein translocation across the endoplasmic reticulum. I. Detection in the microsomal membrane of a receptor for the signal recognition particle. J Cell Biol. 1982 Nov;95(2 Pt 1):463–469. [PMC free article] [PubMed]
  • Gilmore R, Walter P, Blobel G. Protein translocation across the endoplasmic reticulum. II. Isolation and characterization of the signal recognition particle receptor. J Cell Biol. 1982 Nov;95(2 Pt 1):470–477. [PMC free article] [PubMed]
  • Hauser S, Bacher G, Dobberstein B, Lütcke H. A complex of the signal sequence binding protein and the SRP RNA promotes translocation of nascent proteins. EMBO J. 1995 Nov 15;14(22):5485–5493. [PMC free article] [PubMed]
  • Lanzetta PA, Alvarez LJ, Reinach PS, Candia OA. An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem. 1979 Nov 15;100(1):95–97. [PubMed]
  • Lauring B, Sakai H, Kreibich G, Wiedmann M. Nascent polypeptide-associated complex protein prevents mistargeting of nascent chains to the endoplasmic reticulum. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5411–5415. [PMC free article] [PubMed]
  • Luirink J, ten Hagen-Jongman CM, van der Weijden CC, Oudega B, High S, Dobberstein B, Kusters R. An alternative protein targeting pathway in Escherichia coli: studies on the role of FtsY. EMBO J. 1994 May 15;13(10):2289–2296. [PMC free article] [PubMed]
  • Macfarlane J, Müller M. The functional integration of a polytopic membrane protein of Escherichia coli is dependent on the bacterial signal-recognition particle. Eur J Biochem. 1995 Nov 1;233(3):766–771. [PubMed]
  • Meyer DI, Krause E, Dobberstein B. Secretory protein translocation across membranes-the role of the "docking protein'. Nature. 1982 Jun 24;297(5868):647–650. [PubMed]
  • Miller JD, Bernstein HD, Walter P. Interaction of E. coli Ffh/4.5S ribonucleoprotein and FtsY mimics that of mammalian signal recognition particle and its receptor. Nature. 1994 Feb 17;367(6464):657–659. [PubMed]
  • Montoya G, Svensson C, Luirink J, Sinning I. Crystal structure of the NG domain from the signal-recognition particle receptor FtsY. Nature. 1997 Jan 23;385(6614):365–368. [PubMed]
  • Ng DT, Brown JD, Walter P. Signal sequences specify the targeting route to the endoplasmic reticulum membrane. J Cell Biol. 1996 Jul;134(2):269–278. [PMC free article] [PubMed]
  • Phillips GJ, Silhavy TJ. The E. coli ffh gene is necessary for viability and efficient protein export. Nature. 1992 Oct 22;359(6397):744–746. [PubMed]
  • Poritz MA, Bernstein HD, Strub K, Zopf D, Wilhelm H, Walter P. An E. coli ribonucleoprotein containing 4.5S RNA resembles mammalian signal recognition particle. Science. 1990 Nov 23;250(4984):1111–1117. [PubMed]
  • Powers T, Walter P. Reciprocal stimulation of GTP hydrolysis by two directly interacting GTPases. Science. 1995 Sep 8;269(5229):1422–1424. [PubMed]
  • Powers T, Walter P. The nascent polypeptide-associated complex modulates interactions between the signal recognition particle and the ribosome. Curr Biol. 1996 Mar 1;6(3):331–338. [PubMed]
  • Rapiejko PJ, Gilmore R. Protein translocation across the ER requires a functional GTP binding site in the alpha subunit of the signal recognition particle receptor. J Cell Biol. 1992 May;117(3):493–503. [PMC free article] [PubMed]
  • Ribes V, Römisch K, Giner A, Dobberstein B, Tollervey D. E. coli 4.5S RNA is part of a ribonucleoprotein particle that has properties related to signal recognition particle. Cell. 1990 Nov 2;63(3):591–600. [PubMed]
  • Römisch K, Webb J, Herz J, Prehn S, Frank R, Vingron M, Dobberstein B. Homology of 54K protein of signal-recognition particle, docking protein and two E. coli proteins with putative GTP-binding domains. Nature. 1989 Aug 10;340(6233):478–482. [PubMed]
  • Seluanov A, Bibi E. FtsY, the prokaryotic signal recognition particle receptor homologue, is essential for biogenesis of membrane proteins. J Biol Chem. 1997 Jan 24;272(4):2053–2055. [PubMed]
  • Siegel V, Walter P. Elongation arrest is not a prerequisite for secretory protein translocation across the microsomal membrane. J Cell Biol. 1985 Jun;100(6):1913–1921. [PMC free article] [PubMed]
  • Strub K, Moss J, Walter P. Binding sites of the 9- and 14-kilodalton heterodimeric protein subunit of the signal recognition particle (SRP) are contained exclusively in the Alu domain of SRP RNA and contain a sequence motif that is conserved in evolution. Mol Cell Biol. 1991 Aug;11(8):3949–3959. [PMC free article] [PubMed]
  • Tajima S, Lauffer L, Rath VL, Walter P. The signal recognition particle receptor is a complex that contains two distinct polypeptide chains. J Cell Biol. 1986 Oct;103(4):1167–1178. [PMC free article] [PubMed]
  • Ulbrandt ND, Newitt JA, Bernstein HD. The E. coli signal recognition particle is required for the insertion of a subset of inner membrane proteins. Cell. 1997 Jan 24;88(2):187–196. [PubMed]
  • Valent QA, Kendall DA, High S, Kusters R, Oudega B, Luirink J. Early events in preprotein recognition in E. coli: interaction of SRP and trigger factor with nascent polypeptides. EMBO J. 1995 Nov 15;14(22):5494–5505. [PMC free article] [PubMed]
  • Walter P, Blobel G. Translocation of proteins across the endoplasmic reticulum III. Signal recognition protein (SRP) causes signal sequence-dependent and site-specific arrest of chain elongation that is released by microsomal membranes. J Cell Biol. 1981 Nov;91(2 Pt 1):557–561. [PMC free article] [PubMed]
  • Walter P, Blobel G. Preparation of microsomal membranes for cotranslational protein translocation. Methods Enzymol. 1983;96:84–93. [PubMed]
  • Walter P, Johnson AE. Signal sequence recognition and protein targeting to the endoplasmic reticulum membrane. Annu Rev Cell Biol. 1994;10:87–119. [PubMed]
  • Walter P, Jackson RC, Marcus MM, Lingappa VR, Blobel G. Tryptic dissection and reconstitution of translocation activity for nascent presecretory proteins across microsomal membranes. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1795–1799. [PMC free article] [PubMed]
  • Walter P, Ibrahimi I, Blobel G. Translocation of proteins across the endoplasmic reticulum. I. Signal recognition protein (SRP) binds to in-vitro-assembled polysomes synthesizing secretory protein. J Cell Biol. 1981 Nov;91(2 Pt 1):545–550. [PMC free article] [PubMed]
  • Wickner W, Driessen AJ, Hartl FU. The enzymology of protein translocation across the Escherichia coli plasma membrane. Annu Rev Biochem. 1991;60:101–124. [PubMed]
  • Young JC, Ursini J, Legate KR, Miller JD, Walter P, Andrews DW. An amino-terminal domain containing hydrophobic and hydrophilic sequences binds the signal recognition particle receptor alpha subunit to the beta subunit on the endoplasmic reticulum membrane. J Biol Chem. 1995 Jun 30;270(26):15650–15657. [PubMed]
  • Zelazny A, Seluanov A, Cooper A, Bibi E. The NG domain of the prokaryotic signal recognition particle receptor, FtsY, is fully functional when fused to an unrelated integral membrane polypeptide. Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6025–6029. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...