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. 1988 Mar 1;106(3):545–556. doi: 10.1083/jcb.106.3.545

A 5S rRNA/L5 complex is a precursor to ribosome assembly in mammalian cells

PMCID: PMC2115095  PMID: 3279045

Abstract

A novel 5S RNA-protein (RNP) complex in human and mouse cells has been analyzed using patient autoantibodies. The RNP is small (approximately 7S) and contains most of the nonribosome-associated 5S RNA molecules in HeLa cells. The 5S RNA in the particle is matured at its 3' end, consistent with the results of in vivo pulse-chase experiments which indicate that this RNP represents a later step in 5S biogenesis than a previously described 5S*/La protein complex. The protein moiety of the 5S RNP has been identified as ribosomal protein L5, which is known to be released from ribosomes in a complex with 5S after various treatments of the 60S subunit. Indirect immunofluorescence indicates that the L5/5S complex is concentrated in the nucleolus. L5 may therefore play a role in delivering 5S rRNA to the nucleolus for assembly into ribosomes.

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Selected References

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  1. Blobel G. Isolation of a 5S RNA-protein complex from mammalian ribosomes. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1881–1885. doi: 10.1073/pnas.68.8.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blobel G., Sabatini D. Dissociation of mammalian polyribosomes into subunits by puromycin. Proc Natl Acad Sci U S A. 1971 Feb;68(2):390–394. doi: 10.1073/pnas.68.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Branch A. D., Benenfeld B. J., Robertson H. D. Ultraviolet light-induced crosslinking reveals a unique region of local tertiary structure in potato spindle tuber viroid and HeLa 5S RNA. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6590–6594. doi: 10.1073/pnas.82.19.6590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brow D. A., Geiduschek E. P. Modulation of yeast 5 S rRNA synthesis in vitro by ribosomal protein YL3. A possible regulatory loop. J Biol Chem. 1987 Oct 15;262(29):13953–13958. [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Engelke D. R., Ng S. Y., Shastry B. S., Roeder R. G. Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes. Cell. 1980 Mar;19(3):717–728. doi: 10.1016/s0092-8674(80)80048-1. [DOI] [PubMed] [Google Scholar]
  8. Hardin J. A., Rahn D. R., Shen C., Lerner M. R., Wolin S. L., Rosa M. D., Steitz J. A. Antibodies from patients with connective tissue diseases bind specific subsets of cellular RNA-protein particles. J Clin Invest. 1982 Jul;70(1):141–147. doi: 10.1172/JCI110587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hendrick J. P., Wolin S. L., Rinke J., Lerner M. R., Steitz J. A. Ro small cytoplasmic ribonucleoproteins are a subclass of La ribonucleoproteins: further characterization of the Ro and La small ribonucleoproteins from uninfected mammalian cells. Mol Cell Biol. 1981 Dec;1(12):1138–1149. doi: 10.1128/mcb.1.12.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huber P. W., Wool I. G. Use of the cytotoxic nuclease alpha-sarcin to identify the binding site on eukaryotic 5 S ribosomal ribonucleic acid for the ribosomal protein L5. J Biol Chem. 1986 Mar 5;261(7):3002–3005. [PubMed] [Google Scholar]
  11. Keese P., Symons R. H. Domains in viroids: evidence of intermolecular RNA rearrangements and their contribution to viroid evolution. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4582–4586. doi: 10.1073/pnas.82.14.4582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kloetzel P. M., Whitfield W., Sommerville J. Analysis of reconstruction of an RNP particle which stores 5S RNA and tRNA in amphibian oocytes. Nucleic Acids Res. 1981 Feb 11;9(3):605–621. doi: 10.1093/nar/9.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knight E., Jr, Darnell J. E. Distribution of 5 s RNA in HeLa cells. J Mol Biol. 1967 Sep 28;28(3):491–502. doi: 10.1016/s0022-2836(67)80099-8. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lastick S. M., McConkey E. H. Exchange and stability of HeLa ribosomal proteins in vivo. J Biol Chem. 1976 May 25;251(10):2867–2875. [PubMed] [Google Scholar]
  16. Lebleu B., Marbaix G., Huez G., Temmerman J., Burny A., Chantrenne H. Characterization of the messenger ribonucleoprotein released from reticulocyte polyribosomes by EDTA treatment. Eur J Biochem. 1971 Mar 11;19(2):264–269. doi: 10.1111/j.1432-1033.1971.tb01313.x. [DOI] [PubMed] [Google Scholar]
  17. Leibowitz R. D., Weinberg R. A., Penman S. Unusual metabolism of 5 S RNA in HeLa cells. J Mol Biol. 1973 Jan;73(1):139–144. doi: 10.1016/0022-2836(73)90166-6. [DOI] [PubMed] [Google Scholar]
  18. Lerner E. A., Lerner M. R., Janeway C. A., Jr, Steitz J. A. Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease. Proc Natl Acad Sci U S A. 1981 May;78(5):2737–2741. doi: 10.1073/pnas.78.5.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lerner M. R., Boyle J. A., Hardin J. A., Steitz J. A. Two novel classes of small ribonucleoproteins detected by antibodies associated with lupus erythematosus. Science. 1981 Jan 23;211(4480):400–402. doi: 10.1126/science.6164096. [DOI] [PubMed] [Google Scholar]
  20. Lerner M. R., Boyle J. A., Mount S. M., Wolin S. L., Steitz J. A. Are snRNPs involved in splicing? Nature. 1980 Jan 10;283(5743):220–224. doi: 10.1038/283220a0. [DOI] [PubMed] [Google Scholar]
  21. Lin A., Chan Y. L., McNally J., Peleg D., Meyuhas O., Wool I. G. The primary structure of rat ribosomal protein L7. The presence near the amino terminus of L7 of five tandem repeats of a sequence of 12 amino acids. J Biol Chem. 1987 Sep 15;262(26):12665–12671. [PubMed] [Google Scholar]
  22. Lo A. C., Nazar R. N. Topography of 5.8 S rRNA in rat liver ribosomes. Identification of diethyl pyrocarbonate-reactive sites. J Biol Chem. 1982 Apr 10;257(7):3516–3524. [PubMed] [Google Scholar]
  23. Long E. O., Dawid I. B. Repeated genes in eukaryotes. Annu Rev Biochem. 1980;49:727–764. doi: 10.1146/annurev.bi.49.070180.003455. [DOI] [PubMed] [Google Scholar]
  24. Marion M. J., Reboud J. P. An argument for the existence of a natural complex between protein L5 and 5 S RNA in rat liver 60-S ribosomal subunits. Biochim Biophys Acta. 1981 Jan 29;652(1):193–203. doi: 10.1016/0005-2787(81)90222-7. [DOI] [PubMed] [Google Scholar]
  25. Mattaj I. W., Coppard N. J., Brown R. S., Clark B. F., De Robertis E. M. 42S p48--the most abundant protein in previtellogenic Xenopus oocytes--resembles elongation factor 1 alpha structurally and functionally. EMBO J. 1987 Aug;6(8):2409–2413. doi: 10.1002/j.1460-2075.1987.tb02519.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  27. Nazar R. N., Wildeman A. G. Three helical domains form a protein binding site in the 5S RNA-protein complex from eukaryotic ribosomes. Nucleic Acids Res. 1983 May 25;11(10):3155–3168. doi: 10.1093/nar/11.10.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nazar R. N., Yaguchi M., Willick G. E., Rollin C. F., Roy C. The 5-S RNA binding protein from yeast (Saccharomyces cerevisiae) ribosomes. Evolution of the eukaryotic 5-S RNA binding protein. Eur J Biochem. 1979 Dec 17;102(2):573–582. doi: 10.1111/j.1432-1033.1979.tb04274.x. [DOI] [PubMed] [Google Scholar]
  29. Pelham H. R., Brown D. D. A specific transcription factor that can bind either the 5S RNA gene or 5S RNA. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4170–4174. doi: 10.1073/pnas.77.7.4170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Petermann M. L., Hamilton M. G., Pavlovec A. A 5S ribonucleic acid-protein complex extracted from rat liver ribosomes by formamide. Biochemistry. 1972 Jun 6;11(12):2323–2326. doi: 10.1021/bi00762a018. [DOI] [PubMed] [Google Scholar]
  31. Phillips W. F., McConkey E. H. Relative stoichiometry in ribosomal proteins in HeLa cell nucleoli. J Biol Chem. 1976 May 25;251(10):2876–2881. [PubMed] [Google Scholar]
  32. Picard B., Wegnez M. Isolation of a 7S particle from Xenopus laevis oocytes: a 5S RNA-protein complex. Proc Natl Acad Sci U S A. 1979 Jan;76(1):241–245. doi: 10.1073/pnas.76.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Picard B., le Maire M., Wegnez M., Denis H. Biochemical Research on oogenesis. Composition of the 42-S storage particles of Xenopus laevix oocytes. Eur J Biochem. 1980 Aug;109(2):359–368. doi: 10.1111/j.1432-1033.1980.tb04802.x. [DOI] [PubMed] [Google Scholar]
  34. Rinke J., Steitz J. A. Precursor molecules of both human 5S ribosomal RNA and transfer RNAs are bound by a cellular protein reactive with anti-La lupus antibodies. Cell. 1982 May;29(1):149–159. doi: 10.1016/0092-8674(82)90099-x. [DOI] [PubMed] [Google Scholar]
  35. Rosa M. D., Hendrick J. P., Jr, Lerner M. R., Steitz J. A., Reichlin M. A mammalian tRNAHis-containing antigen is recognized by the polymyositis-specific antibody anti-Jo-1. Nucleic Acids Res. 1983 Feb 11;11(3):853–870. doi: 10.1093/nar/11.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schumacher J., Sänger H. L., Riesner D. Subcellular localization of viroids in highly purified nuclei from tomato leaf tissue. EMBO J. 1983;2(9):1549–1555. doi: 10.1002/j.1460-2075.1983.tb01622.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stefano J. E. Purified lupus antigen La recognizes an oligouridylate stretch common to the 3' termini of RNA polymerase III transcripts. Cell. 1984 Jan;36(1):145–154. doi: 10.1016/0092-8674(84)90083-7. [DOI] [PubMed] [Google Scholar]
  38. Terao K., Takahashi Y., Ogata K. Differences between the protein moieties of active subunits and EDTA-treated subunits of rat liver ribosomes with specific references to a 5 S rRNA - protein complex. Biochim Biophys Acta. 1975 Aug 21;402(2):230–237. doi: 10.1016/0005-2787(75)90042-8. [DOI] [PubMed] [Google Scholar]
  39. Ulbrich N., Wool I. G. Identification by affinity chromatography of the eukaryotic ribosomal proteins that bind to 5 S ribosomal ribonucleic acid. J Biol Chem. 1978 Dec 25;253(24):9049–9052. [PubMed] [Google Scholar]
  40. Warner J. R., Soeiro R. Nascent ribosomes from HeLa cells. Proc Natl Acad Sci U S A. 1967 Nov;58(5):1984–1990. doi: 10.1073/pnas.58.5.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wolin S. L., Steitz J. A. The Ro small cytoplasmic ribonucleoproteins: identification of the antigenic protein and its binding site on the Ro RNAs. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1996–2000. doi: 10.1073/pnas.81.7.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

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