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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Oct 15;88(20):9193–9197. doi: 10.1073/pnas.88.20.9193

Site-specific cleavage by metal ion cofactors and inhibitors of M1 RNA, the catalytic subunit of RNase P from Escherichia coli.

S Kazakov 1, S Altman 1
PMCID: PMC52679  PMID: 1718000

Abstract

The location of phosphate residues involved in specific centers for binding of metal ions in M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli, was determined by analysis of induction of cleavage of RNA by metal ions. At pH 9.5, Mg2+ catalyzes cleavage of M1 RNA at five principal sites. Under certain conditions, Mn2+ and Ca2+ can each replace Mg2+ as the cofactor in the processing of precursor tRNAs by M1 RNA and P RNA, the RNA subunit of RNase P from Bacillus subtilis. These cations, as well as various metal ion inhibitors of the catalytic activity of M1 RNA, also promote cleavage of M1 RNA in a specific manner. Certain conditions that affect the catalytic activity of M1 RNA also alter the rate of metal ion-induced cleavage at the various sites. From these results and a comparison of cleavage of M1 RNA with that of a deletion mutant of M1 RNA and of P RNA, we have identified two different centers for binding of metal ions in M1 RNA that are important for the processing of the precursor to tRNA(Tyr) from E. coli. There is also a center for the binding of metal ions in the substrate, close to the site of cleavage by M1 RNA.

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Selected References

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  1. Bartkiewicz M., Gold H., Altman S. Identification and characterization of an RNA molecule that copurifies with RNase P activity from HeLa cells. Genes Dev. 1989 Apr;3(4):488–499. doi: 10.1101/gad.3.4.488. [DOI] [PubMed] [Google Scholar]
  2. Beese L. S., Steitz T. A. Structural basis for the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I: a two metal ion mechanism. EMBO J. 1991 Jan;10(1):25–33. doi: 10.1002/j.1460-2075.1991.tb07917.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown J. W., Haas E. S., James B. D., Hunt D. A., Liu J. S., Pace N. R. Phylogenetic analysis and evolution of RNase P RNA in proteobacteria. J Bacteriol. 1991 Jun;173(12):3855–3863. doi: 10.1128/jb.173.12.3855-3863.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown R. S., Dewan J. C., Klug A. Crystallographic and biochemical investigation of the lead(II)-catalyzed hydrolysis of yeast phenylalanine tRNA. Biochemistry. 1985 Aug 27;24(18):4785–4801. doi: 10.1021/bi00339a012. [DOI] [PubMed] [Google Scholar]
  5. Butzow J. J., Eichhorn G. L. Interactions of metal ions with polynucleotides and related compounds. IV. Degradation of polyribonucleotides by zinc and other divalent metal ions. Biopolymers. 1965;3(1):95–107. doi: 10.1002/bip.360030110. [DOI] [PubMed] [Google Scholar]
  6. Ciesiołka J., Wrzesinski J., Górnicki P., Podkowiński J., Krzyzosiak W. J. Analysis of magnesium, europium and lead binding sites in methionine initiator and elongator tRNAs by specific metal-ion-induced cleavages. Eur J Biochem. 1989 Dec 8;186(1-2):71–77. doi: 10.1111/j.1432-1033.1989.tb15179.x. [DOI] [PubMed] [Google Scholar]
  7. Gardiner K. J., Marsh T. L., Pace N. R. Ion dependence of the Bacillus subtilis RNase P reaction. J Biol Chem. 1985 May 10;260(9):5415–5419. [PubMed] [Google Scholar]
  8. Grosshans C. A., Cech T. R. Metal ion requirements for sequence-specific endoribonuclease activity of the Tetrahymena ribozyme. Biochemistry. 1989 Aug 22;28(17):6888–6894. doi: 10.1021/bi00443a017. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. Guerrier-Takada C., Lumelsky N., Altman S. Specific interactions in RNA enzyme-substrate complexes. Science. 1989 Dec 22;246(4937):1578–1584. doi: 10.1126/science.2480641. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Jack A., Ladner J. E., Rhodes D., Brown R. S., Klug A. A crystallographic study of metal-binding to yeast phenylalanine transfer RNA. J Mol Biol. 1977 Apr 15;111(3):315–328. doi: 10.1016/s0022-2836(77)80054-5. [DOI] [PubMed] [Google Scholar]
  15. Kazakov S. A., Astashkina T. G., Mamaev S. V., Vlassov V. V. Site-specific cleavage of single-stranded DNAs at unique sites by a copper-dependent redox reaction. Nature. 1988 Sep 8;335(6186):186–188. doi: 10.1038/335186a0. [DOI] [PubMed] [Google Scholar]
  16. Kim E. E., Wyckoff H. W. Reaction mechanism of alkaline phosphatase based on crystal structures. Two-metal ion catalysis. J Mol Biol. 1991 Mar 20;218(2):449–464. doi: 10.1016/0022-2836(91)90724-k. [DOI] [PubMed] [Google Scholar]
  17. Lawrence N., Wesolowski D., Gold H., Bartkiewicz M., Guerrier-Takada C., McClain W. H., Altman S. Characteristics of ribonuclease P from various organisms. Cold Spring Harb Symp Quant Biol. 1987;52:233–238. doi: 10.1101/sqb.1987.052.01.028. [DOI] [PubMed] [Google Scholar]
  18. Perreault J. P., Labuda D., Usman N., Yang J. H., Cedergren R. Relationship between 2'-hydroxyls and magnesium binding in the hammerhead RNA domain: a model for ribozyme catalysis. Biochemistry. 1991 Apr 23;30(16):4020–4025. doi: 10.1021/bi00230a029. [DOI] [PubMed] [Google Scholar]
  19. Pyle A. M., McSwiggen J. A., Cech T. R. Direct measurement of oligonucleotide substrate binding to wild-type and mutant ribozymes from Tetrahymena. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8187–8191. doi: 10.1073/pnas.87.21.8187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rordorf B. F., Kearns D. R. Effects of europium (III) on the thermal denaturation and cleavage of transfer ribonucleic acids. Biopolymers. 1976 Aug;15(8):1491–1504. doi: 10.1002/bip.1976.360150805. [DOI] [PubMed] [Google Scholar]
  21. Rubin J. R., Wang J., Sundaralingam M. X-ray diffraction study of the zinc(II) binding sites in yeast phenylalanine transfer RNA. Preferential binding of zinc to guanines in purine-purine sequences. Biochim Biophys Acta. 1983 Mar 15;756(1):111–118. doi: 10.1016/0304-4165(83)90030-2. [DOI] [PubMed] [Google Scholar]
  22. Sugimoto N., Kierzek R., Turner D. H. Kinetics for reaction of a circularized intervening sequence with CU, UCU, CUCU, and CUCUCU: mechanistic implications from the dependence on temperature and on oligomer and Mg2+ concentrations. Biochemistry. 1988 Aug 23;27(17):6384–6392. doi: 10.1021/bi00417a029. [DOI] [PubMed] [Google Scholar]
  23. Surratt C. K., Carter B. J., Payne R. C., Hecht S. M. Metal ion and substrate structure dependence of the processing of tRNA precursors by RNase P and M1 RNA. J Biol Chem. 1990 Dec 25;265(36):22513–22519. [PubMed] [Google Scholar]
  24. Vary C. P., Vournakis J. N. RNA structure analysis using methidiumpropyl-EDTA.Fe(II): a base-pair-specific RNA structure probe. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6978–6982. doi: 10.1073/pnas.81.22.6978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wintermeyer W., Zachau H. G. Mg 2+ -katalysierte, spezifische Spaltung von tRN. Biochim Biophys Acta. 1973 Feb 23;299(1):82–90. [PubMed] [Google Scholar]
  26. Zaug A. J., Kent J. R., Cech T. R. Reactions of the intervening sequence of the Tetrahymena ribosomal ribonucleic acid precursor: pH dependence of cyclization and site-specific hydrolysis. Biochemistry. 1985 Oct 22;24(22):6211–6218. doi: 10.1021/bi00343a027. [DOI] [PubMed] [Google Scholar]

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