Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Apr 11;16(7):2931–2942. doi: 10.1093/nar/16.7.2931

Dependence of M1 RNA substrate specificity on magnesium ion concentration.

L Nichols 1, F J Schmidt 1
PMCID: PMC336442  PMID: 2453026

Abstract

We have constructed a plasmid expressing E. coli M1 RNA, the catalytic RNA subunit of ribonuclease P, under the control of a phage T7 promoter. The active M1 RNA species synthesized in vitro by T7 RNA polymerase from this vector was reacted with the tRNA(Gln) - tRNA(Leu) precursor RNA (Band K) encoded by phage T4. Only the tRNA(Leu) moiety of this dimeric precursor RNA contains the 3' terminal C-C-A sequence common to all tRNAs. We observed that protein-free M1 RNA was capable of processing the precursor RNA at the 5' ends of both tRNA tRNA sequences. The rate of cleavage of the tRNA(Gln) sequence was more strongly dependent on [Mg2+] than that of tRNA(Leu), increasing severalfold between 100 and 500 mM Mg2+, conditions under which the rate of cleavage at the tRNA(Leu) sequence was constant.

Full text

PDF
2931

Images in this article

2934Image
on p.2934
2935Image
on p.2935
2936Image
on p.2936
2937Image
on p.2937
2938Image
on p.2938

Selected References

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

  1. Been M. D., Cech T. R. One binding site determines sequence specificity of Tetrahymena pre-rRNA self-splicing, trans-splicing, and RNA enzyme activity. Cell. 1986 Oct 24;47(2):207–216. doi: 10.1016/0092-8674(86)90443-5. [DOI] [PubMed] [Google Scholar]
  2. Cech T. R. The chemistry of self-splicing RNA and RNA enzymes. Science. 1987 Jun 19;236(4808):1532–1539. doi: 10.1126/science.2438771. [DOI] [PubMed] [Google Scholar]
  3. Guerrier-Takada C., Altman S. M1 RNA with large terminal deletions retains its catalytic activity. Cell. 1986 Apr 25;45(2):177–183. doi: 10.1016/0092-8674(86)90381-8. [DOI] [PubMed] [Google Scholar]
  4. Guerrier-Takada C., Altman S. Structure in solution of M1 RNA, the catalytic subunit of ribonuclease P from Escherichia coli. Biochemistry. 1984 Dec 18;23(26):6327–6334. doi: 10.1021/bi00321a006. [DOI] [PubMed] [Google Scholar]
  5. Guerrier-Takada C., Gardiner K., Marsh T., Pace N., Altman S. The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme. Cell. 1983 Dec;35(3 Pt 2):849–857. doi: 10.1016/0092-8674(83)90117-4. [DOI] [PubMed] [Google Scholar]
  6. Guerrier-Takada C., Haydock K., Allen L., Altman S. Metal ion requirements and other aspects of the reaction catalyzed by M1 RNA, the RNA subunit of ribonuclease P from Escherichia coli. Biochemistry. 1986 Apr 8;25(7):1509–1515. doi: 10.1021/bi00355a006. [DOI] [PubMed] [Google Scholar]
  7. Guerrier-Takada C., McClain W. H., Altman S. Cleavage of tRNA precursors by the RNA subunit of E. coli ribonuclease P (M1 RNA) is influenced by 3'-proximal CCA in the substrates. Cell. 1984 Aug;38(1):219–224. doi: 10.1016/0092-8674(84)90543-9. [DOI] [PubMed] [Google Scholar]
  8. Guthrie C. The nucleotide sequence of the dimeric precursor to glutamine and leucine transfer RNAs coded by bacteriophage T4. J Mol Biol. 1975 Jul 15;95(4):529–547. doi: 10.1016/0022-2836(75)90315-0. [DOI] [PubMed] [Google Scholar]
  9. Hansen F. G., Hansen E. B., Atlung T. Physical mapping and nucleotide sequence of the rnpA gene that encodes the protein component of ribonuclease P in Escherichia coli. Gene. 1985;38(1-3):85–93. doi: 10.1016/0378-1119(85)90206-9. [DOI] [PubMed] [Google Scholar]
  10. Inoue T., Sullivan F. X., Cech T. R. New reactions of the ribosomal RNA precursor of Tetrahymena and the mechanism of self-splicing. J Mol Biol. 1986 May 5;189(1):143–165. doi: 10.1016/0022-2836(86)90387-6. [DOI] [PubMed] [Google Scholar]
  11. Mead D. A., Szczesna-Skorupa E., Kemper B. Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1986 Oct-Nov;1(1):67–74. doi: 10.1093/protein/1.1.67. [DOI] [PubMed] [Google Scholar]
  12. Motamedi H., Lee K., Nichols L., Schmidt F. J. An RNA species involved in Escherichia coli ribonuclease P activity. Gene cloning and effect on transfer RnA synthesis in vivo. J Mol Biol. 1982 Dec 15;162(3):535–550. doi: 10.1016/0022-2836(82)90387-4. [DOI] [PubMed] [Google Scholar]
  13. Motamedi H., Lee Y., Schmidt F. J. Tandem promoters preceding the gene for the M1 RNA component of Escherichia coli ribonuclease P. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3959–3963. doi: 10.1073/pnas.81.13.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Sakamoto H., Kimura N., Nagawa F., Shimura Y. Nucleotide sequence and stability of the RNA component of RNase P from a temperature-sensitive mutant of E. coli. Nucleic Acids Res. 1983 Dec 10;11(23):8237–8251. doi: 10.1093/nar/11.23.8237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schmidt F. J., McClain W. H. Transfer RNA biosynthesis. Alternate orders of ribonuclease P cleavage occur in vitro but not in vivo. J Biol Chem. 1978 Jul 10;253(13):4730–4738. [PubMed] [Google Scholar]
  17. Schmidt F. J., Seidman J. G., Bock R. M. Transfer ribonucleic acid biosynthesis. Substrate specificity of ribonuclease P. J Biol Chem. 1976 Apr 25;251(8):2440–2445. [PubMed] [Google Scholar]
  18. Seidman J. G., McClain W. H. Three steps in conversion of large precursor RNA into serine and proline transfer RNAs. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1491–1495. doi: 10.1073/pnas.72.4.1491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Seidman J. G., Schmidt F. J., Foss K., McClain W. H. A mutant of escherichia coli defective in removing 3' terminal nucleotides from some transfer RNA precursor molecules. Cell. 1975 Aug;5(4):389–400. doi: 10.1016/0092-8674(75)90058-6. [DOI] [PubMed] [Google Scholar]
  20. Subbarao M. N., Makam H., Apirion D. A site in a tRNA precursor that can be processed by the whole RNase P enzyme but not by the RNA alone. J Biol Chem. 1984 Dec 10;259(23):14339–14342. [PubMed] [Google Scholar]
  21. Wilson J. H., Abelson J. N. Bacteriophage T4 transfer RNA. II. Mutants of T4 defective in the formation of functional suppressor transfer RNA. J Mol Biol. 1972 Aug 14;69(1):57–73. doi: 10.1016/0022-2836(72)90023-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES