Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Mar 25;16(6):2565–2583. doi: 10.1093/nar/16.6.2565

RNA binding proteins of the large subunit of bovine mitochondrial ribosomes.

M A Piatyszek 1, N D Denslow 1, T W O'Brien 1
PMCID: PMC336390  PMID: 3129699

Abstract

RNA binding properties of proteins from the large subunit of bovine mitochondrial ribosomes were studied using four different approaches: binding of radiolabeled RNA to western blotted proteins; disassembly of the intact 39 S ribosomal subunits with urea; binding of ribosomal proteins to RNA in the presence of urea; and binding of proteins extracted with lithium chloride to RNA. Results from these four approaches allowed us to identify a set of six proteins (L7, L13, L14, L21, L26, and L44) which appear to be strong RNA binding proteins. Seven additional proteins (L8, L11, L28, L35, L40, L49, and L50) were identified as secondary RNA binding proteins. RNA binding properties of the proteins in both of these sets were compared with the topographic disposition and susceptibility towards lithium chloride extraction of the individual proteins. Proteins from the first set are good candidates for early assembly proteins since they have a high affinity for RNA, are generally found in 4M lithium chloride core particles, and are among the most buried proteins in the 39 S subunit.

Full text

PDF
2565

Images in this article

2571Image
on p.2571
2572Image
on p.2572
2573Image
on p.2573
2574Image
on p.2574
2577Image
on p.2577

Selected References

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

  1. Anderson S., Bankier A. T., Barrell B. G., de Bruijn M. H., Coulson A. R., Drouin J., Eperon I. C., Nierlich D. P., Roe B. A., Sanger F. Sequence and organization of the human mitochondrial genome. Nature. 1981 Apr 9;290(5806):457–465. doi: 10.1038/290457a0. [DOI] [PubMed] [Google Scholar]
  2. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. doi: 10.1093/nar/8.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brimacombe R., Stöffler G., Wittmann H. G. Ribosome structure. Annu Rev Biochem. 1978;47:217–249. doi: 10.1146/annurev.bi.47.070178.001245. [DOI] [PubMed] [Google Scholar]
  4. Cohlberg J. A. Conditions for ethanol precipitation of active 50 S ribosomal subunits from Escherichia coli. Anal Biochem. 1980 Jul 15;106(1):195–198. doi: 10.1016/0003-2697(80)90137-2. [DOI] [PubMed] [Google Scholar]
  5. Denslow N. D., O'Brien T. W. Antibiotic susceptibility of the peptidyl transferase locus of bovine mitochondrial ribosomes. Eur J Biochem. 1978 Nov 15;91(2):441–448. doi: 10.1111/j.1432-1033.1978.tb12696.x. [DOI] [PubMed] [Google Scholar]
  6. Denslow N. D., O'Brien T. W. Elongation factors EF-G from E. coli and mammalian mitochondria are not functionally interchangeable. Biochem Biophys Res Commun. 1979 Oct 29;90(4):1257–1265. doi: 10.1016/0006-291x(79)91172-0. [DOI] [PubMed] [Google Scholar]
  7. Denslow N. D., O'Brien T. W. Organization of proteins in mammalian mitochondrial ribosomes. Accessibility to lactoperoxidase-catalyzed radioiodination. J Biol Chem. 1984 Aug 10;259(15):9867–9873. [PubMed] [Google Scholar]
  8. Dijk J., Littlechild J., Garrett R. A. The RNA binding properties of "native" protein-protein complexes isolated from the Escherichia coli ribosome. FEBS Lett. 1977 May 15;77(2):295–300. doi: 10.1016/0014-5793(77)80255-x. [DOI] [PubMed] [Google Scholar]
  9. Dijk J., Littlechild J. Purification of ribosomal proteins from Escherichia coli under nondenaturing conditions. Methods Enzymol. 1979;59:481–502. doi: 10.1016/0076-6879(79)59109-5. [DOI] [PubMed] [Google Scholar]
  10. El-Baradi T. T., Raué H. A., De Regt C. H., Planta R. J. Stepwise dissociation of yeast 60S ribosomal subunits by LiCl and identification of L25 as a primary 26S rRNA binding protein. Eur J Biochem. 1984 Oct 15;144(2):393–400. doi: 10.1111/j.1432-1033.1984.tb08477.x. [DOI] [PubMed] [Google Scholar]
  11. Giri L., Dijk J. Physical studies on proteins L3 and L24 from the 50 S subunit of the Escherichia coli ribosome. Arch Biochem Biophys. 1979 Mar;193(1):122–129. doi: 10.1016/0003-9861(79)90015-8. [DOI] [PubMed] [Google Scholar]
  12. Gourse R. L., Thurlow D. L., Gerbi S. A., Zimmermann R. A. Specific binding of a prokaryotic ribosomal protein to a eukaryotic ribosomal RNA: implications for evolution and autoregulation. Proc Natl Acad Sci U S A. 1981 May;78(5):2722–2726. doi: 10.1073/pnas.78.5.2722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hamilton M. G., O'Brien T. W. Ultracentrifugal characterization of the mitochondrial ribosome and subribosomal particles of bovine liver: molecular size and composition. Biochemistry. 1974 Dec 17;13(26):5400–5403. doi: 10.1021/bi00723a024. [DOI] [PubMed] [Google Scholar]
  14. Hardy S. J., Kurland C. G., Voynow P., Mora G. The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochemistry. 1969 Jul;8(7):2897–2905. doi: 10.1021/bi00835a031. [DOI] [PubMed] [Google Scholar]
  15. Matthews D. E., Hessler R. A., Denslow N. D., Edwards J. S., O'Brien T. W. Protein composition of the bovine mitochondrial ribosome. J Biol Chem. 1982 Aug 10;257(15):8788–8794. [PubMed] [Google Scholar]
  16. Mougel M., Ehresmann B., Ehresmann C. Binding of Escherichia coli ribosomal protein S8 to 16S rRNA: kinetic and thermodynamic characterization. Biochemistry. 1986 May 20;25(10):2756–2765. doi: 10.1021/bi00358a003. [DOI] [PubMed] [Google Scholar]
  17. Nierhaus K. H., Dohme F. Total reconstitution of 50 S subunits from Escherichia coli ribosomes. Methods Enzymol. 1979;59:443–449. doi: 10.1016/0076-6879(79)59106-x. [DOI] [PubMed] [Google Scholar]
  18. Noller H. F. Structure of ribosomal RNA. Annu Rev Biochem. 1984;53:119–162. doi: 10.1146/annurev.bi.53.070184.001003. [DOI] [PubMed] [Google Scholar]
  19. O'Brien T. W. The general occurrence of 55 S ribosomes in mammalian liver mitochondria. J Biol Chem. 1971 May 25;246(10):3409–3417. [PubMed] [Google Scholar]
  20. Rozier C., Mache R. Binding of 16S rRNA to chloroplast 30S ribosomal proteins blotted on nitrocellulose. Nucleic Acids Res. 1984 Oct 11;12(19):7293–7304. doi: 10.1093/nar/12.19.7293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Salacinski P. R., McLean C., Sykes J. E., Clement-Jones V. V., Lowry P. J. Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenyl glycoluril (Iodogen). Anal Biochem. 1981 Oct;117(1):136–146. doi: 10.1016/0003-2697(81)90703-x. [DOI] [PubMed] [Google Scholar]
  22. Schieber G. L., O'Brien T. W. Extraction of proteins from the large subunit of bovine mitochondrial ribosomes under nondenaturing conditions. J Biol Chem. 1982 Aug 10;257(15):8781–8787. [PubMed] [Google Scholar]
  23. Schieber G. L., O'Brien T. W. Site of synthesis of the proteins of mammalian mitochondrial ribosomes. Evidence from cultured bovine cells. J Biol Chem. 1985 May 25;260(10):6367–6372. [PubMed] [Google Scholar]
  24. Traub P., Nomura M. Structure and function of E. coli ribosomes. V. Reconstitution of functionally active 30S ribosomal particles from RNA and proteins. Proc Natl Acad Sci U S A. 1968 Mar;59(3):777–784. doi: 10.1073/pnas.59.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wolff P., Gilz R., Schumacher J., Riesner D. Complexes of viroids with histones and other proteins. Nucleic Acids Res. 1985 Jan 25;13(2):355–367. doi: 10.1093/nar/13.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Woodcock-Mitchell J., Eichner R., Nelson W. G., Sun T. T. Immunolocalization of keratin polypeptides in human epidermis using monoclonal antibodies. J Cell Biol. 1982 Nov;95(2 Pt 1):580–588. doi: 10.1083/jcb.95.2.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wrede P., Erdmann V. A. Escherichia coli 5S RNA binding proteins L18 and L25 interact with 5.8S RNA but not with 5S RNA from yeast ribosomes. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2706–2709. doi: 10.1073/pnas.74.7.2706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. el-Baradi T. T., Raué H. A., de Regt V. C., Verbree E. C., Planta R. J. Yeast ribosomal protein L25 binds to an evolutionary conserved site on yeast 26S and E. coli 23S rRNA. EMBO J. 1985 Aug;4(8):2101–2107. doi: 10.1002/j.1460-2075.1985.tb03898.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES