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. 1978 Jan;133(1):251–255. doi: 10.1128/jb.133.1.251-255.1978

Effects of Detergents on Ribosomal Precursor Subunits of Bacillus megaterium

Barbara A Body 1,†, Bernard H Brownstein 1,††
PMCID: PMC222001  PMID: 412833

Abstract

Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction.

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Selected References

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

  1. BRITTEN R. J., McCARTHY B. J., ROBERTS R. B. The synthesis of ribosomes in E. coli. IV. The synthesis of ribosomal protein and the assembly of ribosomes. Biophys J. 1962 Jan;2:83–93. doi: 10.1016/s0006-3495(62)86842-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Body B. A., Brownstein B. H. Ribosomal precursor particles of Bacillus megaterium. J Bacteriol. 1976 Jun;126(3):1149–1155. doi: 10.1128/jb.126.3.1149-1155.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bryant R. E., Sypherd P. S. Genetic analysis of cold-sensitive ribosome maturation mutants of Escherichia coli. J Bacteriol. 1974 Mar;117(3):1082–1092. doi: 10.1128/jb.117.3.1082-1092.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Di Girolamo M., Hinckley E., Busiello E. Localization of robosome precursors in Escherichia coli. Biochim Biophys Acta. 1968 Dec 17;169(2):387–397. doi: 10.1016/0005-2787(68)90047-6. [DOI] [PubMed] [Google Scholar]
  5. Dimmitt K., Sypherd P. S. Association of lysozyme with ribosomes and ribosome precursor particles. J Bacteriol. 1973 Nov;116(2):1059–1061. doi: 10.1128/jb.116.2.1059-1061.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guthrie C., Nashimoto H., Nomura M. Structure and function of E. coli ribosomes. 8. Cold-sensitive mutants defective in ribosome assembly. Proc Natl Acad Sci U S A. 1969 Jun;63(2):384–391. doi: 10.1073/pnas.63.2.384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hayes F., Hayes D. H. Biosynthesis of ribosomes in E. coli. I. Properties of ribosomal precursor particles and their RNA components. Biochimie. 1971;53(3):369–382. doi: 10.1016/s0300-9084(71)80104-9. [DOI] [PubMed] [Google Scholar]
  8. Haywood A. M. Cellular site of Escherichia coli ribosomal RNA synthesis. Proc Natl Acad Sci U S A. 1971 Feb;68(2):435–439. doi: 10.1073/pnas.68.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lester R. L., Steiner M. R. The occurrence of diphosphoinositide and triphosphoinositide in Saccharomyces cerevisiae. J Biol Chem. 1968 Sep 25;243(18):4889–4893. [PubMed] [Google Scholar]
  10. Lewandowski L. J., Brownstein B. L. Characterization of a 43 s ribonucleoprotein component of a mutant of Escherichia coli. J Mol Biol. 1969 Apr;41(2):277–290. doi: 10.1016/0022-2836(69)90392-1. [DOI] [PubMed] [Google Scholar]
  11. Mangiarotti G., Apirion D., Schlessinger D., Silengo L. Biosynthetic precursors of 30S and 50S ribosomal particles in Escherichia coli. Biochemistry. 1968 Jan;7(1):456–472. doi: 10.1021/bi00841a058. [DOI] [PubMed] [Google Scholar]
  12. Mangiarotti G., Turco E., Perlo C., Altruda F. Role of precursor 16S RNA in assembly of E. coli 30S ribosomes. Nature. 1975 Feb 13;253(5492):569–571. doi: 10.1038/253569a0. [DOI] [PubMed] [Google Scholar]
  13. Mangiarotti G., Turco E., Ponzetto A., Altruda F. Precursor 16S RNA in active 30S ribosomes. Nature. 1974 Jan 18;247(5437):147–148. doi: 10.1038/247147a0. [DOI] [PubMed] [Google Scholar]
  14. McCarthy B. J., Britten R. J., Roberts R. B. The Synthesis of Ribosomes in E. coli: III. Synthesis of Ribosomal RNA. Biophys J. 1962 Jan;2(1):57–82. doi: 10.1016/s0006-3495(62)86841-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Osawa S., Otaka E., Itoh T., Fukui T. Biosynthesis of 50 s ribosomal subunit in Escherichia coli. J Mol Biol. 1969 Mar 28;40(3):321–351. doi: 10.1016/0022-2836(69)90158-2. [DOI] [PubMed] [Google Scholar]
  16. Roth G. S., Daneo-Moore L. Intracellular localization of ribosome biosynthesis in osmotically fragile forms of Streptococcus faecalis. Biochim Biophys Acta. 1971 Jul 29;240(4):575–587. doi: 10.1016/0005-2787(71)90715-5. [DOI] [PubMed] [Google Scholar]
  17. Sypherd P. S., Bryant R., Dimmitt K., Fujisawa T. Genetic control of ribosome assembly. J Supramol Struct. 1974;2(2-4):166–177. doi: 10.1002/jss.400020211. [DOI] [PubMed] [Google Scholar]
  18. Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
  19. van Dijk-Salkinoja M. S., Planta R. J. Formation and life cycle of ribosomal subunits, mono- and polyribosomes in Bacillus licheniformis. Arch Biochem Biophys. 1970 Dec;141(2):477–488. doi: 10.1016/0003-9861(70)90165-7. [DOI] [PubMed] [Google Scholar]
  20. van Dijk-Salkinoja M. S., Stoof T. J., Planta R. J. The distribution of polysomes, ribosomes and ribosomal subunits in exponential-phase cells of Bacillus licheniformis. Eur J Biochem. 1970 Feb;12(3):474–482. doi: 10.1111/j.1432-1033.1970.tb00875.x. [DOI] [PubMed] [Google Scholar]

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