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. 1976 May;3(5):1351–1371. doi: 10.1093/nar/3.5.1351

Cleavage of T4 species I ribonucleic acid by Escherichia coli ribonuclease III.

G V Paddock, K Fukada, J Abelson, H D Robertson
PMCID: PMC342991  PMID: 781626

Abstract

T4 Species I RNA, a molecule 140 nucleotides in length with some structural features very much like a tRNA, is specifically cleaved by an enzymatic activity in Escherichia coli extracts to give three segments with 19, 48 and 73 nucleotides. We report the purification and characterization of the E. coli RNase which cleaves two 3' phosphodiester bonds of T4 Species I RNA. This reaction has many properties in common with those catalyzed by E. coli RNase III, although the optimal salt conditions for T4 Species I RNA cleavage differ significantly from those for other RNase III-catalyzed reactions. The reaction is not catalyzed by extracts from an E. coli strain lacking RNase III activity. Furthermore, T4 Species I RNA is cleaved by highly purified E. coli RNase III to yield the same three specific fragments. We conclude that this specific cleavage is due to the action of RNase III, and that the requirement for lower ionic strength may reveal further important properties about this RNA processing enzyme.

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Selected References

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

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dunn J. J., Studier F. W. T7 early RNAs and Escherichia coli ribosomal RNAs are cut from large precursor RNAs in vivo by ribonuclease 3. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3296–3300. doi: 10.1073/pnas.70.12.3296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dunn J. J., Studier F. W. T7 early RNAs are generated by site-specific cleavages. Proc Natl Acad Sci U S A. 1973 May;70(5):1559–1563. doi: 10.1073/pnas.70.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kindler P., Keil T. U., Hofschneider P. H. Isolation and characterization of a ribonuclease 3 deficient mutant of Escherichia coli. Mol Gen Genet. 1973 Oct 16;126(1):53–59. doi: 10.1007/BF00333481. [DOI] [PubMed] [Google Scholar]
  5. Kramer R. A., Rosenberg M., Steitz J. A. Nucleotide sequences of the 5' and 3' termini of bacteriophage T7 early messenger RNAs synthesized in vivo: evidence for sequence specificity in RNA processing. J Mol Biol. 1974 Nov 15;89(4):767–776. doi: 10.1016/0022-2836(74)90051-5. [DOI] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Paddock G. V., Abelson J. Nucleotide sequence determination of bacteriophage T2 and T6 species I ribonucleic acids. J Biol Chem. 1975 Jun 10;250(11):4207–4219. [PubMed] [Google Scholar]
  8. Paddock G. V., Abelson J. Nucleotide sequence determination of bacteriophage T4 species I ribonucleic acid. J Biol Chem. 1975 Jun 10;250(11):4185–4206. [PubMed] [Google Scholar]
  9. Paddock G., Abelson J. Sequence of T4, T2 and T6 bacteriophage species I RNA and specific cleavage by an E. coli endonuclease. Nat New Biol. 1973 Nov 7;246(149):2–6. doi: 10.1038/newbio246002a0. [DOI] [PubMed] [Google Scholar]
  10. Reiner A. M. Isolation and mapping of polynucleotide phosphorylase mutants of Escherichia coli. J Bacteriol. 1969 Mar;97(3):1431–1436. doi: 10.1128/jb.97.3.1431-1436.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Robertson H. D., Altman S., Smith J. D. Purification and properties of a specific Escherichia coli ribonuclease which cleaves a tyrosine transfer ribonucleic acid presursor. J Biol Chem. 1972 Aug 25;247(16):5243–5251. [PubMed] [Google Scholar]
  12. Robertson H. D., Dunn J. J. Ribonucleic acid processing activity of Escherichia coli ribonuclease III. J Biol Chem. 1975 Apr 25;250(8):3050–3056. [PubMed] [Google Scholar]
  13. Robertson H. D., Webster R. E., Zinder N. D. Purification and properties of ribonuclease III from Escherichia coli. J Biol Chem. 1968 Jan 10;243(1):82–91. [PubMed] [Google Scholar]
  14. Rosenberg M., Kramer R. A., Steitz J. A. T7 early messenger RNAs are the direct products of ribonuclease III cleavage. J Mol Biol. 1974 Nov 15;89(4):777–782. doi: 10.1016/0022-2836(74)90052-7. [DOI] [PubMed] [Google Scholar]
  15. Siegelman H. W., Wieczorek G. A., Turner B. C. Preparation of calcium phosphate for protein chromatography. Anal Biochem. 1965 Dec;13(3):402–404. doi: 10.1016/0003-2697(65)90332-5. [DOI] [PubMed] [Google Scholar]
  16. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  17. Tinoco I., Jr, Uhlenbeck O. C., Levine M. D. Estimation of secondary structure in ribonucleic acids. Nature. 1971 Apr 9;230(5293):362–367. doi: 10.1038/230362a0. [DOI] [PubMed] [Google Scholar]

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