Skip to main content
SpringerLink
Log in

Structural and functional characterisation of the signal recognition particle-specific 54 kDa protein (SRP54) of tomato

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

Two representative genes for the 54 kDa protein subunit of the signal recognition particle (SRP54) of tomato were cloned. It was shown that both genes are expressed in the tomato cv. Rentita. SRP54 is encoded by nine exons distributed over 10 kb of genomic sequence. The amino acid sequences deduced for the two SRP54 genes are 92% identical and the calculated protein size is 55 kDa. Like the homologous proteins isolated from other eukaryotes, the tomato SRP54 is evidently divided into two domains. As deduced from sequence motif identity, the N-terminally located G-domain can be assumed to have GTPase activity. The C-terminal part of the protein is methionine rich (14% methionine) and represents the M-domain. In in vitro binding experiments, SRP54 of tomato was able to attach to the 7S RNA of tomato, its natural binding partner in the SRP. This interaction can only take place in a trimeric complex consisting of 7S RNA, SRP54 and SRP19. The latter protein subunit of the SRP complex is assumed to induce a conformational change in the 7S RNA. The human SRP19 was able to mediate the binding of the tomato SRP54 to the 7S RNA, irrespective of whether this latter originated from tomato or man.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andreazzoli M, Gerbi SA (1991) Changes in the 7SL RNA conformation during the signal recognition particle cycle. EMBO J 10:767–777

    Google Scholar 

  • Bassüner R, Wobus U, Rapoport TA (1984) Signal recognition particle triggers the translocation of storage globulin polypeptides from field beans (Vicia faba L.) across mammalian endoplasmic reticulum membrane. FEBS Lett 166:314–320

    Google Scholar 

  • Bernstein HD, Poritz MA, Strub K, Hoben PJ, Brenner S, Walter P (1989) Model for signal sequence recognition from amino-acid sequence of 54K subunit of signal recognition particle. Nature 340:482–486

    Google Scholar 

  • Brown JWS (1986) A catalogue of splice junctions and putative branch point sequences from plant introns. Nucleic Acids Res 14:9549–9559

    Google Scholar 

  • Byström AS, Hjalmarsson KJ, Wilkström PM, Björk GR (1983) The nucleotide sequence of an Escherichia coli operon containing genes for the tRNA (m1G) methyltransferase, the ribosomal proteins S16 and L19 and a 21-K polypeptide. EMBO J 2:899–905

    Google Scholar 

  • Campos N, Palau J, Torrent M, Ludevid D (1988) Signal recognition-like particles are present in maize. J Biol Chem 263:9646–9650

    Google Scholar 

  • Campos N, Palau J, Zwieb C (1989) Diversity of 7SL RNA from signal recognition particle of maize endosperm. Nucleic Acids Res 17:1573–1588

    Google Scholar 

  • Chamberlain JP (1979) Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem 98:132–135

    Google Scholar 

  • Connolly T, Gilmore R (1989) The signal recognition particle receptor mediates the GTP-dependent displacement of SRP from the nascent polypeptide. Cell 57:599–610

    Google Scholar 

  • Dever TE, Glynias MJ, Merrick WC (1987) GTP-binding domain: three consensus sequence elements with distinct spacing. Proc Natl Acad Sci USA 84:1814–1818

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Franklin AE, Hoffman NE (1993) Characterization of a chloroplast homologue of the 54-kDa subunit of the signal recognition particle. J Biol Chem 268:22175–22180

    Google Scholar 

  • Haas B, Klanner A, Ramm K, Sänger HL (1988) The 7S RNA from tomato leaf tissue resembles a signal recognition RNA and exhibits a remarkable sequence complementarity to viroids. EMBO J 7:4063–4074

    Google Scholar 

  • Hann BC, Poritz MA, Walter P (1989) Saccharomyces cerevisiae and Schizosaccharomyces pombe contain a homologue to the 54 kD subunit of the signal recognition particle that in S. cerevisiae is essential for growth. J Cell Biol 109:3223–3230

    Google Scholar 

  • Higgins TJV, Spencer D (1981) Precursor forms of pea vicilin subunits. Modification by microsomal membranes during cell-free translation. Plant Physiol 67:205–211

    Google Scholar 

  • High S, Dobberstein B (1991) The signal sequence interacts with the methionine-rich domain of the 54 kD protein of signal recognition particle. J Cell Biol 113:229–233

    Google Scholar 

  • Honda K, Nakamura K, Nishiguchi M, Yamane K (1993) Cloning and characterization of a Bacillus subtilis gene encoding a homolog of the 54-kilodalton subunit of mammalian signal recognition particle and Escherichia coli Ffh. J Bacteriol 175:4885–4894

    Google Scholar 

  • Kozak M (1984) Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res 12:857–872

    Google Scholar 

  • Kozak M (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292

    Google Scholar 

  • Krieg UC, Walter P, Johnson AE (1986) Photocrosslinking of the signal sequence of nascent preprolactin to the 54-kilodalton polypeptide of the signal recognition particle. Proc Natl Acad Sci USA 83:8604–8608

    Google Scholar 

  • Kurzchalia TV, Wiedmann M, Girshovich AS, Bochkareva ES, Bielka H, Rapoport TA (1986) The signal sequence of nascent preprolactin interacts with the 54K polypeptide of the signal recognition particle. Nature 320:634–636

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Langridge P, Pintor-Toro JA, Feix G (1982) Zein precursor mRNAs from maize endosperm. Mol Gen Genet 187:432–438

    Google Scholar 

  • Larsen N, Zwieb C (1991) SRP-RNA sequence alignment and secondary structure. Nucleic Acids Res 19:209–215

    Google Scholar 

  • Lauffer L, Garcia PD, Harkins RN, Coussens L, Ullrich A, Walter P (1985) Topology of signal recognition particle receptor in endoplasmic reticulum membrane. Nature 318:334–338

    Google Scholar 

  • Liao X, Selinger D, Althoff S, Chiang A, Hamilton D, Ma M, Wise JA (1992) Random mutagenesis of Schizosaccharomyces pombe SRP RNA: lethal and conditional lesions cluster in presumptive protein binding sites. Nucleic Acids Res 20:1607–1615

    Google Scholar 

  • Lindstrom JT, Chu B, Belanger FC (1993) Isolation and characterization of an Arabidopsis thaliana gene for the 54 kDa subunit of the signal recognition particle. Plant Mol Biol 23:1265–1272

    Google Scholar 

  • Lingelbach K, Zwieb C, Webb JR, Marshallsay C, Hoben PJ, Walter P, Dobberstein B (1988) Isolation and characterization of a cDNA clone encoding the 19 kDa protein of signal recognition particle (SRP): expression and binding to 7SL RNA. Nucleic Acids Res 16:9431–9442

    Google Scholar 

  • Lütcke H, High S, Römisch K, Ashford AJ, Dobberstein B (1992) The methionine-rich domain of the 54 kDa subunit of signal recognition particle is sufficient for the interaction with signal sequences. EMBO J 11:1543–1551

    Google Scholar 

  • Marshallsay C, Prehn S, Zwieb C (1989) cDNA cloning of the wheat germ SRP 7S RNAs. Nucleic Acids 17:1771

    Google Scholar 

  • Miller JD, Wilhelm H, Gierasch L, Gilmore R, Walter P (1993) GTP binding and hydrolysis by the signal recognition particle during initiation of protein translocation. Nature 366:351–354

    Google Scholar 

  • Ogg SC, Nunnari JM, Miller JD, Walter P (1992) The role of GTP in the protein targeting to the endoplasmic reticulum. In: Neupert W, Lill R (eds) Membrane biogenesis and protein targeting. Neuberger A, van Deenen LLM (eds) New comprehensive biochemistry, vol 22. Elsevier/Amsterdam, London, New York, Tokyo, pp 129–136

    Google Scholar 

  • Prehn S, Wiedmann M, Rapoport TA, Zwieb C (1987) Protein translocation across wheat germ microsomal membranes requires an SRP-like component. EMBO J 6:2093–2097

    Google Scholar 

  • Proudfoot NJ, Brownlee GG (1976) 3′ non-coding region sequences in eukaryotic messenger RNA. Nature 263:211–214

    Google Scholar 

  • Riedel L (1993) Analyse der Sekundärstruktur und Charakterisierung der Genfamilie der 7SL (SRP) RNA der Tomate (Lycopersicon esculentum). Dissertation, University of Cologne, Germany

    Google Scholar 

  • Römisch K, Webb J, Herz J, Prehn S, Frank R, Vingron M, Dobberstein B (1989) Homology of 54K protein of signal recognition particle, docking protein and two E. coli proteins with putative GTP-binding domains. Nature 340:478–482

    Google Scholar 

  • Römisch K, Webb J, Lingelbach K, Gausepohl H, Dobberstein B (1990) The 54-kD protein of the signal recognition particle contains a methionine-rich RNA binding domain. J Cell Biol 111:1793–1802

    Google Scholar 

  • Sambrook J, Fritsch EF, Mamatis T (1989) Molecular cloning. A laboratory manual (2nd edn) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Samuelsson T (1992) A Mycoplasma protein homologous to mammalian SRP54 recognizes a highly conserved domain of SRP RNA. Nucleic Acids Res 20:5763–5770

    Google Scholar 

  • Schwarz-Sommer Z, Gierl A, Klösgen RB, Wienand U, Peterson PA, Saedler H (1984) The Spm (En) transposable element controls the excision of a 2 kb DNA insert at the wx m − 8 allele of Zea mays. EMBO J 3:1021–1028

    Google Scholar 

  • Siegel V, Walter P (1988) Each of the activities of signal recognition particle (SRP) is contained within a distinct domain: analysis of biochemical mutants of SRP. Cell 52:39–49

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Strub K, Moss J, Walter P (1991) 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 11:3949–3959

    Google Scholar 

  • Walter P, Blobel G (1980) Purification of a membrane-associated protein complex required for protein translocation across the endoplasmic reticulum. Proc Natl Acad Sci USA 77:7112–7116

    Google Scholar 

  • Walter P, Blobel G (1981) 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 91:557–561

    Google Scholar 

  • Walter P, Blobel G (1982) Signal recognition particle contains a 7S RNA essential for protein translocation across the endoplasmic reticulum. Nature 299:691–698

    Google Scholar 

  • Walter P, Blobel G (1983) Disassembly and reconstitution of signal recognition particle. Cell 34:525–533

    Google Scholar 

  • Woo SLC (1979) A sensitive and rapid method for recombinant plaque screening. Methods Enzymol 68:389–395

    Google Scholar 

  • Zopf D, Bernstein HD, Johnson AE, Walter P (1990) The methionine-rich domain of the 54 kd protein subunit of the signal recognition particle contains an RNA binding site and can be crosslinked to a signal sequence. EMBO J 9:4511–4517

    Google Scholar 

  • Zopf D, Bernstein HD, Walter P (1993) GTPase domain of the 54-kD subunit of the mammalian signal recognition particle is required for protein translocation but not for signal sequence binding. J Cell Biol 120:1131–1121

    Google Scholar 

  • Zwieb C (1992) Recognition of a tetranucleotide loop of signal recognition particle RNA by SRP19. J Biol Chem 267:15650–15656

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Abteilung für Viroidforschung, Max-Planck-Institut für Biochemie, D-82152, Martinsried, Germany

    Sylvia Krolkiewicz, Heinz L. Sänger & Ursula Niesbach-KLösgen

Authors
  1. Sylvia Krolkiewicz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Heinz L. Sänger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ursula Niesbach-KLösgen
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by R. Herrmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Krolkiewicz, S., Sänger, H.L. & Niesbach-KLösgen, U. Structural and functional characterisation of the signal recognition particle-specific 54 kDa protein (SRP54) of tomato. Molec. Gen. Genet. 245, 565–576 (1994). https://doi.org/10.1007/BF00282219

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00282219

Key words

Search

Navigation