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. 1987 Jan 1;104(1):61–66. doi: 10.1083/jcb.104.1.61

Signal recognition particle causes a transient arrest in the biosynthesis of prepromelittin and mediates its translocation across mammalian endoplasmic reticulum

PMCID: PMC2117042  PMID: 3025225

Abstract

The translocation of prepromelittin (pPM) across mammalian endoplasmic reticulum was studied in both wheat germ and reticulocyte lysate. In the wheat germ system, signal recognition particle (SRP) caused a transient arrest in the synthesis of pPM. This was indicated by a slowdown in the rate of synthesis of pPM in the presence of SRP. The arrest was specific, dependent on the concentration of SRP, and more effective at early incubation time. In a tightly synchronized translation system, SRP had no apparent effect on the elongation of pPM, indicating that the effect of SRP on pPM chain synthesis might be at the final stages of chain elongation and release from the ribosome. This was reflected in a transient accumulation of pPM as peptidyl tRNA. Because pPM is composed of only 70 amino acids, arrest by SRP may be very close to chain termination. Arrest at this stage of chain synthesis seems to be unstable and the nascent chain gets terminated and released from the ribosome after a transient delay. The translocation of pPM was shown to be dependent on both SRP and docking protein. The difference in the translocation efficiency of pPM in reticulocyte and wheat germ lysates may reflect a difference in the targeting process in the two systems.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ainger K. J., Meyer D. I. Translocation of nascent secretory proteins across membranes can occur late in translation. EMBO J. 1986 May;5(5):951–955. doi: 10.1002/j.1460-2075.1986.tb04308.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blobel G., Dobberstein B. Transfer of proteins across membranes. II. Reconstitution of functional rough microsomes from heterologous components. J Cell Biol. 1975 Dec;67(3):852–862. doi: 10.1083/jcb.67.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans E. A., Gilmore R., Blobel G. Purification of microsomal signal peptidase as a complex. Proc Natl Acad Sci U S A. 1986 Feb;83(3):581–585. doi: 10.1073/pnas.83.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Friedlander M., Blobel G. Bovine opsin has more than one signal sequence. 1985 Nov 28-Dec 4Nature. 318(6044):338–343. doi: 10.1038/318338a0. [DOI] [PubMed] [Google Scholar]
  6. Gilmore R., Blobel G. Transient involvement of signal recognition particle and its receptor in the microsomal membrane prior to protein translocation. Cell. 1983 Dec;35(3 Pt 2):677–685. doi: 10.1016/0092-8674(83)90100-9. [DOI] [PubMed] [Google Scholar]
  7. Gilmore R., Blobel G. Translocation of secretory proteins across the microsomal membrane occurs through an environment accessible to aqueous perturbants. Cell. 1985 Sep;42(2):497–505. doi: 10.1016/0092-8674(85)90107-2. [DOI] [PubMed] [Google Scholar]
  8. 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. doi: 10.1083/jcb.95.2.463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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. doi: 10.1083/jcb.95.2.470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haeuptle M. T., Frank R., Dobberstein B. Translation arrest by oligodeoxynucleotides complementary to mRNA coding sequences yields polypeptides of predetermined length. Nucleic Acids Res. 1986 Feb 11;14(3):1427–1448. doi: 10.1093/nar/14.3.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hobden A. N., Cundliffe E. The mode of action of alpha sarcin and a novel assay of the puromycin reaction. Biochem J. 1978 Jan 15;170(1):57–61. doi: 10.1042/bj1700057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hortsch M., Meyer D. I. Transfer of secretory proteins through the membrane of the endoplasmic reticulum. Int Rev Cytol. 1986;102:215–242. doi: 10.1016/s0074-7696(08)61276-0. [DOI] [PubMed] [Google Scholar]
  13. Ibrahimi I. M., Cutler D., Stueber D., Bujard H. Determinants for protein translocation across mammalian endoplasmic reticulum. Membrane insertion of truncated and full-length prelysozyme molecules. Eur J Biochem. 1986 Mar 17;155(3):571–576. doi: 10.1111/j.1432-1033.1986.tb09526.x. [DOI] [PubMed] [Google Scholar]
  14. Kaschnitz R., Kreil G. Processing of prepromelittin by subcellular fractions from rat liver. Biochem Biophys Res Commun. 1978 Aug 14;83(3):901–907. doi: 10.1016/0006-291x(78)91480-8. [DOI] [PubMed] [Google Scholar]
  15. Kindas-Mügge I., Lane C. D., Kreil G. Insect protein synthesis in frog cells: the translation of honey bee promelittin messenger RNA in Xenopus oocytes. J Mol Biol. 1974 Aug 15;87(3):451–462. doi: 10.1016/0022-2836(74)90096-5. [DOI] [PubMed] [Google Scholar]
  16. Levin I. W., Lavialle F., Mollay C. Comparative effects of melittin and its hydrophobic and hydrophilic fragments on bilayer organization by Raman spectroscopy. Biophys J. 1982 Jan;37(1):339–349. doi: 10.1016/S0006-3495(82)84682-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meyer D. I. Signal recognition particle (SRP) does not mediate a translational arrest of nascent secretory proteins in mammalian cell-free systems. EMBO J. 1985 Aug;4(8):2031–2033. doi: 10.1002/j.1460-2075.1985.tb03888.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mollay C., Kreil G., Berger H. Action of phospholipases on the cytoplasmic membrane of Escherichia coli. Stimulation by melittin. Biochim Biophys Acta. 1976 Mar 5;426(2):317–324. doi: 10.1016/0005-2736(76)90340-0. [DOI] [PubMed] [Google Scholar]
  19. Mollay C., Vilas U., Kreil G. Cleavage of honeybee prepromelittin by an endoprotease from rat liver microsomes: identification of intact signal peptide. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2260–2263. doi: 10.1073/pnas.79.7.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Posch M., Rakusch U., Mollay C., Laggner P. Cooperative effects in the interaction between melittin and phosphatidylcholine model membranes. Studies by temperature scanning densitometry. J Biol Chem. 1983 Feb 10;258(3):1761–1766. [PubMed] [Google Scholar]
  21. 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. doi: 10.1083/jcb.100.6.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Siegel V., Walter P. Removal of the Alu structural domain from signal recognition particle leaves its protein translocation activity intact. Nature. 1986 Mar 6;320(6057):81–84. doi: 10.1038/320081a0. [DOI] [PubMed] [Google Scholar]
  23. Suchanek G., Kreil G., Hermodson M. A. Amino acid sequence of honeybee prepromelittin synthesized in vitro. Proc Natl Acad Sci U S A. 1978 Feb;75(2):701–704. doi: 10.1073/pnas.75.2.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Suchanek G., Kreil G. Translation of melittin messenger RNA in vitro yields a product terminating with glutaminylglycine rather than with glutaminamide. Proc Natl Acad Sci U S A. 1977 Mar;74(3):975–978. doi: 10.1073/pnas.74.3.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Walter P., Blobel G. Purification of a membrane-associated protein complex required for protein translocation across the endoplasmic reticulum. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7112–7116. doi: 10.1073/pnas.77.12.7112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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. doi: 10.1083/jcb.91.2.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Walter P., Gilmore R., Blobel G. Protein translocation across the endoplasmic reticulum. Cell. 1984 Aug;38(1):5–8. doi: 10.1016/0092-8674(84)90520-8. [DOI] [PubMed] [Google Scholar]
  28. 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. doi: 10.1083/jcb.91.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wickner W. T., Lodish H. F. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. doi: 10.1126/science.4048938. [DOI] [PubMed] [Google Scholar]
  30. Zimmermann R., Mollay C. Import of honeybee prepromelittin into the endoplasmic reticulum. Requirements for membrane insertion, processing, and sequestration. J Biol Chem. 1986 Sep 25;261(27):12889–12895. [PubMed] [Google Scholar]

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