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. 1972 Mar;109(3):1284–1294. doi: 10.1128/jb.109.3.1284-1294.1972

“Compartmentalization” of Escherichia coli Ribosomes and Ribonucleic Acid

Frederick Varricchio 1
PMCID: PMC247353  PMID: 4551751

Abstract

Escherichia coli spheroplasts lysed by Brij 58 and deoxycholate were separated into supernatant (S) and membrane fractions by low-speed centrifugation. The membrane fraction was further divided into that which was releasable by deoxyribonuclease (fraction D) and that which was not (M). In the presence of 10−2m Mg2+, the S, D, and M fractions contained, respectively, 60, 20, and 20% of the total cellular ribonucleic acid (RNA). Ribosomal and transfer RNA (rRNA, tRNA) were found in each fraction. The M + D fraction RNA was labeled more by a pulse label. Incorporation of uracil into the D fraction continued only as long as the uptake of exogenous uracil, suggesting that this was a major primary site of RNA synthesis. From pulse-labeled cells, each fraction contained precursor rRNA, and there was a 10S RNA in the M fraction. Ninety per cent of the ribosomal subunits and the ribosomal precursor particles, 26 and 43S, were in the S fraction. Precursor RNA (17S) was found in the 26S precursor particles. The D fraction contained 38% of the polysomes (this does not consider polysomes, if any, of the M fraction) which were labeled four times as much as the supernatant polysomes by a 1-min pulse of uracil. These results are interpreted to mean that new RNA is associated with a cytoplasmic membrane-RNA polymerase-DNA complex.

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Selected References

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  1. Adesnik M., Levinthal C. RNA metabolism in T4-infected Escherichia coli. J Mol Biol. 1970 Mar 14;48(2):187–208. doi: 10.1016/0022-2836(70)90156-7. [DOI] [PubMed] [Google Scholar]
  2. Bulova S. I., Burka E. R. Biosynthesis of nonglobin protein by membrane-bound ribosomes in reticulocytes. J Biol Chem. 1970 Oct 10;245(19):4907–4912. [PubMed] [Google Scholar]
  3. CHAMBERLIN M., BERG P. Deoxyribo ucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1962 Jan 15;48:81–94. doi: 10.1073/pnas.48.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cundliffe E. Intracellular distribution of ribosomes and polyribosomes in Bacillus megaterium. J Mol Biol. 1970 Sep 28;52(3):467–481. doi: 10.1016/0022-2836(70)90413-4. [DOI] [PubMed] [Google Scholar]
  5. Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Fairbanks G., Jr, Levinthal C., Reeder R. H. Analysis of C14-labeled proteins by disc electrophoresis. Biochem Biophys Res Commun. 1965 Aug 16;20(4):393–399. doi: 10.1016/0006-291x(65)90589-9. [DOI] [PubMed] [Google Scholar]
  8. Fuchs E., Hanawalt P. Isolation and characterization of the DNA replication complex from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):301–322. doi: 10.1016/0022-2836(70)90032-x. [DOI] [PubMed] [Google Scholar]
  9. Gesteland R. F. Isolation and characterization of ribonuclease I mutants of Escherichia coli. J Mol Biol. 1966 Mar;16(1):67–84. doi: 10.1016/s0022-2836(66)80263-2. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. KONO M., OSAWA S. INTERMEDIARY STEPS OF RIBOSOME FORMATION IN ESCHERICHIA COLI. Biochim Biophys Acta. 1964 Jun 22;87:326–334. doi: 10.1016/0926-6550(64)90228-2. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. McCarthy B. J., Britten R. J. The Synthesis of Ribosomes in E. coli: I. The Incorporation of C-Uracil into the Metabolic Pool and RNA. Biophys J. 1962 Jan;2(1):35–47. doi: 10.1016/s0006-3495(62)86839-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Patterson D., Weinstein M., Nixon R., Gillespie D. Interaction of ribosomes and the cell envelope of Escherichia coli mediated by lysozyme. J Bacteriol. 1970 Feb;101(2):584–591. doi: 10.1128/jb.101.2.584-591.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  16. Pettijohn D. E., Clarkson K., Kossman C. R., Stonington O. G. Synthesis of ribosomal RNA on a protein-DNA complex isolated from bacteria: a comparison of ribosomal RNA synthesis in vitro and in vivo. J Mol Biol. 1970 Sep 14;52(2):281–300. doi: 10.1016/0022-2836(70)90031-8. [DOI] [PubMed] [Google Scholar]
  17. Phillips L. A., Hotham-Iglewski B., Franklin R. M. Polyribosomes of Escherichia coli. I. Effects of monovalent cations on the distribution of polysomes, ribosomes and ribosomal subunits. J Mol Biol. 1969 Mar 14;40(2):279–288. doi: 10.1016/0022-2836(69)90475-6. [DOI] [PubMed] [Google Scholar]
  18. Rouvière J., Lederberg S., Granboulan P., Gros F. Structural sites of RNA synthesis in Escherichia coli. J Mol Biol. 1969 Dec 28;46(3):413–430. doi: 10.1016/0022-2836(69)90185-5. [DOI] [PubMed] [Google Scholar]
  19. Ryter A. Association of the nucleus and the membrane of bacteria: a morphological study. Bacteriol Rev. 1968 Mar;32(1):39–54. doi: 10.1128/br.32.1.39-54.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. SCHLESSINGER D. PROTEIN SYNTHESIS BY POLYRIBOSOMES ON PROTOPLAST MEMBRANES OF B. MEGATERIUM. J Mol Biol. 1963 Nov;7:569–582. doi: 10.1016/s0022-2836(63)80103-5. [DOI] [PubMed] [Google Scholar]
  21. Takagi M., Tanaka T., Ogata K. Functional differences in protein synthesis between free and bound polysomes of rat liver. Biochim Biophys Acta. 1970 Sep 17;217(1):148–158. doi: 10.1016/0005-2787(70)90131-0. [DOI] [PubMed] [Google Scholar]
  22. Traub P., Nomura M. Structure and function of Escherichia coli ribosomes. VI. Mechanism of assembly of 30 s ribosomes studied in vitro. J Mol Biol. 1969 Mar 28;40(3):391–413. doi: 10.1016/0022-2836(69)90161-2. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Varricchio F., Monier R. Ribosome patterns in Escherichia coli growing at various rates. J Bacteriol. 1971 Oct;108(1):105–110. doi: 10.1128/jb.108.1.105-110.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Varrichio F. Polysomes patterns in E. coli and K12 at various growth rates. Bull Soc Chim Biol (Paris) 1969;51(10):1547–1548. [PubMed] [Google Scholar]
  26. YUDKIN M. D., DAVIS B. NATURE OF THE RNA ASSOCIATED WITH THE PROTOPLAST MEMBRANE OF BACILLUS MEGATERIUM. J Mol Biol. 1965 May;12:193–204. doi: 10.1016/s0022-2836(65)80293-5. [DOI] [PubMed] [Google Scholar]

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