Skip to main content
Log in

Characterization of 10S RNA: A new stable RNA molecule from Escherichia coli

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

When cells of Escherichia coli are labeled with 32Pi for long periods of time and the cell content is subjected to electrophoresis in polyacrylamide gels, an RNA band appears which is about 10S in size. This band seems to contain three conformers. After treatment with formamide only a single band appears in this region of the gel, which contains 550 nucleotides as determined from its mobility. The complexity of the fingerprint of this material, after digestion with T1-RNase, is in agreement with the size as determined by the mobility, this confirming that indeed it is a single molecule. Composition of the T1-oligonucleotides was determined by digesting the T1-generated oligonucleotides with pancreatic RNase and T2-RNase. The quantitative and qualitative analysis of these digestions suggests that 10S RNA contains 609 nucleotides. The molecule contains, besides the four regular bases, one copy per molecule of the modified base pseudouridine.

10S RNA cannot be processed by cell extracts to tRNA-sized molecules and does not bind significantly to ribosomes, hence it is unlikely to be a tRNA precursor or an mRNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bailey, S.C., Apirion, D.: Repetitive DNA in Escherichia coli: multiple sequences complementary to small stable RNAs. Molec. Gen. Genet. 1979 in press

  • Branlant, C., Ebel, J.P.: Studies on the primary structure of E. coli 23S RNA. Nucleotide sequence of the ribonuclease T1 digestion products containing more than one uridine residue. J. Mol. Biol. 111, 215–256 (1977)

    Google Scholar 

  • Brownlee, G.G.: Sequence of 6S RNA of E. coli. Nature New Biol. 229 147–149 (1971)

    Google Scholar 

  • Brownlee, G.G., Sanger, F., Barrell, B.G.: Nucleotide sequence of 5S-ribosomal RNA from E. coli. Nature 215, 735–736 (1967)

    Google Scholar 

  • Brownlee, G.G., Sanger, F., Barrell, B.G.: The sequence of 5S ribosomal ribonucleic acid. J. Mol. Biol. 34, 379–412 (1968)

    Google Scholar 

  • Fellner, P., Ehresmann, C., Ebel, J.P.: The determination of the primary structure of the 16S ribosomal RNA of E. coli 1. Nucleotide sequence analysis of T1 and pancreatic RNase digestion products. Biochimie 54, 853–900 (1972)

    Google Scholar 

  • Gegenheimer, P., Apirion, D.: Processing of rRNA by RNAse P: Spacer tRNAs are linked to 16S rRNA in an RNAase P RNAase III mutant strain of E. coli. Cell 15, 527–539 (1978)

    Google Scholar 

  • Goldstein, J., Harwood, K.: Another species of ribonucleic acid in E. coli. J. Mol. Biol. 39, 383–387 (1969)

    Google Scholar 

  • Griffin, B.E.: Studies and sequences of E. coli 4.5S RNA. J. Biol. Chem. 250, 5426–5437 (1975)

    Google Scholar 

  • Hindley, J.: Fractionation of 32P-labeled ribonucleic acids on polyacrylamide gels and their characterization by fingerprinting. J. Mol. Biol. 30, 125–136 (1967)

    Google Scholar 

  • Ikemura, T., Dahlberg, J.E.: Small ribonucleic acids of E. coli I. Characterization by polyacrylamide gel electrophoresis and fingerprint analysis. J. Biol. Chem. 248, 5024–5032 (1973)

    Google Scholar 

  • Ilgen, C., Kirk, L.L., Carbon, J.: Isolation and characterization of large transfer ribonucleic acid precursors from E. coli. J. Biol. Chem. 251, 922–929 (1976)

    Google Scholar 

  • Kaplan, R., Apirion, D.: The fate of ribosome in E. coli cells starved for a carbon source. J. Biol. Chem. 250, 1854–1863 (1975)

    Google Scholar 

  • Kim, K.T., Apirion, D., Ghora, B.K.: Small stable RNA of Neurospora crassa. Mol. Gen. Genet. 160, 25–32 (1978)

    Google Scholar 

  • Lee, S.Y., Bailey, S.C., Apirion, D.: Small stable RNAs from E. coli: Evidence for the existence of new molecules and for a new ribonucleoprotein particle containing 6S RNA. J. Bacteriol. 133, 1015–1023 (1978)

    Google Scholar 

  • Loening, U.: The determination of the molecular weight of ribonucleic acid by polyacrylamide gel electrophoresis. The effects of changes in conformation. Biochem. J. 113, 131–138 (1969)

    Google Scholar 

  • Saneyoshi, M., Harada, F., Nishimura, S.: Isolation and characterization of N6-methyladenosine from E. coli valine t-RNA. Biochim. Biophys. Acta 190, 264–273 (1969)

    Google Scholar 

  • Sanger, F., Brownlee, G.G., Barrell, B.G.: A two-dimensional fractionation procedure for radioactive nucleotides. J. Mol. Biol. 13, 373–398 (1965)

    Google Scholar 

  • Schedl, P., Primakoff, P.: Mutants of E. coli thermosensitive for the synthesis of transfer RNA. Proc. Natl. Acad. Sci U.S.A. 70, 2091–2095 (1973)

    Google Scholar 

  • Steitz, J.A.: Nucleotide sequences of the ribosomal binding sites of bacteriophage R17 RNA. Cold Spring Harbor Symp. Quant. Biol. 34, 621–633 (1969)

    Google Scholar 

  • Swanstrom, R., Shank, P.R.: X-ray intensifying screens greatly enhance the detection by autoradiography of the radioactive isotopes 32P and 125I. Analyt. Biochem. 86, 184–192 (1978)

    Google Scholar 

  • Uchida, T., Bonen, L., Schaup, H.W., Lewis, B.J., Zablen, L., Woese, C.: The use of ribonuclease U2 in RNA sequence determination. J. Mol. Evolut. 3, 63–77 (1974)

    Google Scholar 

  • Villareal, L.P., Breindl, M., Holland, J.J.: Determination of molar ratios of vesicular stomatitis virus induced RNA species in BHK21 cells. Biochemistry 15, 1663–1667 (1976)

    Google Scholar 

  • Volckaert, G., Fiers, W.: Micro thin-layer techniques for rapid sequence analysis of 32P-labeled RNA: Double digestion and pancreatic ribonuclease analyses. Analyt. Biochem. 83, 228–239 (1977)

    Google Scholar 

  • Volckaert, G., Min Jou, W., Fiers, W.: Analysis of 32P-labeled bacteriophage MS2 RNA by mini-fingerprinting procedure. Analyt. Biochem. 72, 433–446 (1976)

    Google Scholar 

  • Weatherford, S.C., Rosen, L., Gorelic, L., Apirion, D.: E. coli strains with thermolabile ribonuclease II activity. J. Biol. Chem. 247, 5404–5408 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H.G. Wittmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ray, B.K., Apirion, D. Characterization of 10S RNA: A new stable RNA molecule from Escherichia coli . Molec. Gen. Genet. 174, 25–32 (1979). https://doi.org/10.1007/BF00433301

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00433301

Keywords

Navigation