Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Jun 11;19(11):2955–2960. doi: 10.1093/nar/19.11.2955

Interaction of protein SRP19 with signal recognition particle RNA lacking individual RNA-helices.

C Zwieb 1
PMCID: PMC328257  PMID: 1711676

Abstract

Derivatives of human SRP-RNA were constructed by site-directed mutagenesis and tested for their ability to interact with protein SRP19. An RNA missing helix 6 barely interacts with SRP19, while the helix 8-deletion mutant retains much binding capability. A mutant RNA consisting just of helix 6 also binds the protein, but not as well as the unaltered molecule. SRP19 interacts to a full extent with the fourth mutant RNA composed of helices 6, 7, 8 and a portion of helix 5. It is concluded that helix 6- and not helix 8- is the major SRP19 binding site. Helices 7, 8 and portions of helix 5 contribute to the formation of a functional site. These results agree with data suggesting a proximity of helix 6 and the conserved part of SRP-RNA.

Full text

PDF
2955

Images in this article

2958Image
on p.2958
2958Image
on p.2958
2959Image
on p.2959
2959Image
on p.2959

Selected References

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

  1. Andrews D. W., Walter P., Ottensmeyer F. P. Evidence for an extended 7SL RNA structure in the signal recognition particle. EMBO J. 1987 Nov;6(11):3471–3477. doi: 10.1002/j.1460-2075.1987.tb02671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cheong C., Varani G., Tinoco I., Jr Solution structure of an unusually stable RNA hairpin, 5'GGAC(UUCG)GUCC. Nature. 1990 Aug 16;346(6285):680–682. doi: 10.1038/346680a0. [DOI] [PubMed] [Google Scholar]
  3. Gundelfinger E. D., Krause E., Melli M., Dobberstein B. The organization of the 7SL RNA in the signal recognition particle. Nucleic Acids Res. 1983 Nov 11;11(21):7363–7374. doi: 10.1093/nar/11.21.7363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kurzchalia T. V., Wiedmann M., Girshovich A. S., Bochkareva E. S., Bielka H., Rapoport T. A. The signal sequence of nascent preprolactin interacts with the 54K polypeptide of the signal recognition particle. Nature. 1986 Apr 17;320(6063):634–636. doi: 10.1038/320634a0. [DOI] [PubMed] [Google Scholar]
  5. Larsen N., Zwieb C. SRP-RNA sequence alignment and secondary structure. Nucleic Acids Res. 1991 Jan 25;19(2):209–215. doi: 10.1093/nar/19.2.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lingelbach K., Zwieb C., Webb J. R., Marshallsay C., Hoben P. J., Walter P., Dobberstein B. 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. 1988 Oct 25;16(20):9431–9442. doi: 10.1093/nar/16.20.9431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Nelson R. M., Long G. L. A general method of site-specific mutagenesis using a modification of the Thermus aquaticus polymerase chain reaction. Anal Biochem. 1989 Jul;180(1):147–151. doi: 10.1016/0003-2697(89)90103-6. [DOI] [PubMed] [Google Scholar]
  8. Prehn S., Wiedmann M., Rapoport T. A., Zwieb C. Protein translocation across wheat germ microsomal membranes requires an SRP-like component. EMBO J. 1987 Jul;6(7):2093–2097. doi: 10.1002/j.1460-2075.1987.tb02475.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rapoport T. A. Protein transport across the ER membrane. Trends Biochem Sci. 1990 Sep;15(9):355–358. doi: 10.1016/0968-0004(90)90076-n. [DOI] [PubMed] [Google Scholar]
  10. 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. doi: 10.1016/0092-8674(90)90454-m. [DOI] [PubMed] [Google Scholar]
  11. Romaniuk P. J., de Stevenson I. L., Wong H. H. Defining the binding site of Xenopus transcription factor IIIA on 5S RNA using truncated and chimeric 5S RNA molecules. Nucleic Acids Res. 1987 Mar 25;15(6):2737–2755. doi: 10.1093/nar/15.6.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Römisch K., Webb J., Lingelbach K., Gausepohl H., Dobberstein B. The 54-kD protein of signal recognition particle contains a methionine-rich RNA binding domain. J Cell Biol. 1990 Nov;111(5 Pt 1):1793–1802. doi: 10.1083/jcb.111.5.1793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Siegel V., Walter P. Binding sites of the 19-kDa and 68/72-kDa signal recognition particle (SRP) proteins on SRP RNA as determined in protein-RNA "footprinting". Proc Natl Acad Sci U S A. 1988 Mar;85(6):1801–1805. doi: 10.1073/pnas.85.6.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Siegel V., Walter P. Each of the activities of signal recognition particle (SRP) is contained within a distinct domain: analysis of biochemical mutants of SRP. Cell. 1988 Jan 15;52(1):39–49. doi: 10.1016/0092-8674(88)90529-6. [DOI] [PubMed] [Google Scholar]
  15. Toschka H. Y., Struck J. C., Erdmann V. A. The 4.5S RNA gene from Pseudomonas aeruginosa. Nucleic Acids Res. 1989 Jan 11;17(1):31–36. doi: 10.1093/nar/17.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ullu E., Weiner A. M. Human genes and pseudogenes for the 7SL RNA component of signal recognition particle. EMBO J. 1984 Dec 20;3(13):3303–3310. doi: 10.1002/j.1460-2075.1984.tb02294.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Walter P., Blobel G. Disassembly and reconstitution of signal recognition particle. Cell. 1983 Sep;34(2):525–533. doi: 10.1016/0092-8674(83)90385-9. [DOI] [PubMed] [Google Scholar]
  18. Walter P., Lingappa V. R. Mechanism of protein translocation across the endoplasmic reticulum membrane. Annu Rev Cell Biol. 1986;2:499–516. doi: 10.1146/annurev.cb.02.110186.002435. [DOI] [PubMed] [Google Scholar]
  19. Zwieb C., Schüler D. Low resolution three-dimensional models of the 7SL RNA of the signal recognition particle, based on an intramolecular cross-link introduced by mild irradiation with ultraviolet light. Biochem Cell Biol. 1989 Aug;67(8):434–442. doi: 10.1139/o89-069. [DOI] [PubMed] [Google Scholar]
  20. Zwieb C. Structure and function of signal recognition particle RNA. Prog Nucleic Acid Res Mol Biol. 1989;37:207–234. doi: 10.1016/s0079-6603(08)60699-6. [DOI] [PubMed] [Google Scholar]
  21. Zwieb C., Ullu E. Identification of dynamic sequences in the central domain of 7SL RNA. Nucleic Acids Res. 1986 Jun 11;14(11):4639–4657. doi: 10.1093/nar/14.11.4639. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES