Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1975 Apr;2(4):501–507. doi: 10.1093/nar/2.4.501

Modification of E. coli ribosomes and coliphage MS2 RNA by bisulfite: effects on ribosomal binding and protein synthesis.

B Braverman, R Shapiro, W Szer
PMCID: PMC342858  PMID: 1094421

Abstract

The reaction of E. coli 70s ribosomes with 0.2 M NaH-35 s03 (pH 7.1, 3.5hrs, 37 degree) led to the conversion of 4.5% of the uracil residues of the R, RNA into 5.6-dihydrouracil-6-sulfonate residues. The modified ribosomes exhibited a significant decrease in their ability to bind (14-C)-phenylalanyl-(RNA-phe and to incorporate (14-C)-phenylalanine into protein in the presence of polyuridylic acid. The ability of the modified ribosomes to form an initiation complex as measured by the A-U-G or coliphage MS2 RNA dependent binding of (14-C)-fmet-tRNA-fmet was also impaired, as was their ability to incorporate (14-C) lysine into protein with MS2 RNA as messenger. Treatment os MS RNA with 0.2 M sodium (35-S) bisulfite, pH 7.0 at 25 degrees C resulted in the substitution of 2.7% and 6.2% of the uracil residues by bisulfite after 1 and 3.5 hrs of reaction, respectively. Impairment of function of the MS2 RNA in both initiation complex formation and transplantation assays was observed. These reactions of uracil residues of mRNA and rRNA may be a cause of biological damage inflicted by sodium bisulfite and sulfur dioxide.

Full text

PDF
501

Selected References

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

  1. Gunnison A. F., Palmes E. D. Persistence of plasma S-sulfonates following exposure of rabbits to sulfite and sulfur dioxide. Toxicol Appl Pharmacol. 1973 Feb;24(2):266–278. doi: 10.1016/0041-008x(73)90147-6. [DOI] [PubMed] [Google Scholar]
  2. Held W. A., Gette W. R., Nomura M. Role of 16S ribosomal ribonucleic acid and the 30S ribosomal protein S12 in the initiation of natural messenger ribonucleic acid translation. Biochemistry. 1974 May 7;13(10):2115–2122. doi: 10.1021/bi00707a019. [DOI] [PubMed] [Google Scholar]
  3. Leffler S., Szer W. Polypeptide chain initiation in Caulobacter crescentus without initiation factor IF-1. J Biol Chem. 1974 Mar 10;249(5):1465–1468. [PubMed] [Google Scholar]
  4. Leffler S., Szer W. Purification and properties of initiation factor IF-3 from Caulobacter crescentus. J Biol Chem. 1974 Mar 10;249(5):1458–1464. [PubMed] [Google Scholar]
  5. Lodish H. F. Secondary structure of bacteriophage f2 ribonucleic acid and the initiation of in vitro protein biosynthesis. J Mol Biol. 1970 Jun 28;50(3):689–702. doi: 10.1016/0022-2836(70)90093-8. [DOI] [PubMed] [Google Scholar]
  6. Min Jou W., Haegeman G., Ysebaert M., Fiers W. Nucleotide sequence of the gene coding for the bacteriophage MS2 coat protein. Nature. 1972 May 12;237(5350):82–88. doi: 10.1038/237082a0. [DOI] [PubMed] [Google Scholar]
  7. NIRENBERG M., LEDER P. RNA CODEWORDS AND PROTEIN SYNTHESIS. THE EFFECT OF TRINUCLEOTIDES UPON THE BINDING OF SRNA TO RIBOSOMES. Science. 1964 Sep 25;145(3639):1399–1407. doi: 10.1126/science.145.3639.1399. [DOI] [PubMed] [Google Scholar]
  8. STRAUSS J. H., Jr, SINSHEIMER R. L. Purification and properties of bacteriophage MS2 and of its ribonucleic acid. J Mol Biol. 1963 Jul;7:43–54. doi: 10.1016/s0022-2836(63)80017-0. [DOI] [PubMed] [Google Scholar]
  9. Shapiro R., Braverman B., Louis J. B., Servis R. E. Nucleic acid reactivity and conformation. II. Reaction of cytosine and uracil with sodium bisulfite. J Biol Chem. 1973 Jun 10;248(11):4060–4064. [PubMed] [Google Scholar]
  10. Shapiro R., Braverman B. Modification of polyuridylic acid by bisulfite: effect on double helix formation and coding properties. Biochem Biophys Res Commun. 1972 May 12;47(3):544–550. doi: 10.1016/0006-291x(72)90913-8. [DOI] [PubMed] [Google Scholar]
  11. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Stanley W. M., Jr, Bock R. M. Isolation and physical properties of the ribosomal ribonucleic acid of Escherichia coli. Biochemistry. 1965 Jul;4(7):1302–1311. doi: 10.1021/bi00883a014. [DOI] [PubMed] [Google Scholar]
  13. Szer W. Cell-free protein synthesis at 0 degrees. An activating factor from ribosomes of a psychrophilic microorganism. Biochim Biophys Acta. 1970 Jul 16;213(1):159–170. [PubMed] [Google Scholar]
  14. Szer W., Leffler S. Interaction of Escherichia coli 30S ribosomal subunits with MS2 phage RNA in the absence of initiation factors. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3611–3615. doi: 10.1073/pnas.71.9.3611. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES