Effects of Divalent Cations on the Spicing of Phage T4 Thymidylate Synthase Intron RNA

  • Park, In-Kook (Department of Applied Biology, Dongguk University) ;
  • Sung, Jung-Suk (Department of Applied Biology, Dongguk University) ;
  • Shin, Sook (Department of Biology, Sahmyook University)
  • Published : 1997.03.01

Abstract

Effects of divalent cations such as $Mg^{2+}$, $Mn^{2+}$, $Ca^{2+}$, and $Zn^2$ on splicing activity of phage T4 thymidylate synthase intron RNA have been investigated. At the concentration of 0.5 mM, $Mn^{2+}$ in the absence of $Mg^{2+}$, a very small amount of pre-RNA was cleaved into ligation products (El-E2) but no circular or linear intron was produced. As the concentration of $Mn^{2+}$ was increased from 1 to 5 mM the pre-RNA was completely hydrolyzed. In the presence of 5 mM $Mg^{2+}$, both the linear intron and circular intron were produced but no El-E2 ligation product was produced. At both 3 and 5 mM $Mn^{2+}$ the RNA was hydrolyzed completely as observed with no $Mg^2+$ being present. In the case of $Zn^{2+}$, even at 0.5 mM concentration, the pre-RNA was completely hydrolyzed. This observation suggested that $Zn^{2+}$ facilitates RNA hydrolysis more rapidly than $Mn^{2+}$ does. at 5mM $Ca^{2+}$, the RNA was not hydrolyzed and remained intact as a primary transcript.

Keywords

References

  1. Cell v.45 Role of conserved sequence elements 9L and 2 in self-splicing of Tetrahmena ribosomal RNA precursor Burke JM;Irvine KD;Kaneko KJ;Kerker BJ;Oettger AB;Tierney WM;Williamson CL;Zaug AJ;Cech TR
  2. Annu Rev Biochem v.55 Biological catalysis by RNA Cech TR;Bass BL
  3. Science v.236 The chemistry of self-splicing RNA and RNA enzyme Cech TR
  4. science v.251 Visualizing the higher order floding of a catalytic RNA molecule Celander DW;Cech TR
  5. Proc Natl Acad Sci USA v.81 An intervening sequence in the thymidylate synthase gene of bacteriophage T4 Chu FK;Maley GF;Maley F;Belfort M
  6. FASEB J v.2 RNA splicing in the T-even bacteriophage Chu FK;Maley GF;Maley F
  7. Biochemistry v.30 Role of divanlent metalions in the hammerhead RNA cleavage reation Dahm SC;Uhlenbeck OC
  8. Nature v.300 A model for splicing in fungal mitochondria Davies RW;Waring RB;Ray RA;Brown TA;Scazzocchio C
  9. Biochemistry v.28 Metal ion requirements for sequence-specific endonnuclease activity of the Tetrahymena ribozyme Grosshans CA;Cech TR
  10. Biochemistry v.25 Metal ion requirements and other aspects of the reaction catalyzed by M1 RNA subunit of RNase P from E. coli. Guerrier-Takada C;Haydock K;Allen L;Altman S
  11. Nature v.342 The quanosine binding site of the Tetrahymena ribozyme Michel F;Hanna M;Green R;Bartel DP;Szostak JW
  12. Korean Biochem J v.25 The functional role of P2 structure of T4 td intorn in thymidylate synthase activity in vivo ParkIk
  13. Proceedings of the 7th FAOBMB Congress v.2 Effect of monovalent cation and spermidine on RNA splicing of T4 phage td intron ParkIK;SungJS;Shins
  14. Nature v.361 Metal ion catalysis in the Tetraymena ribozyme reaction Piccirlli JA;Vyle JS;Caruthers MH;Cech TR
  15. Science v.261 Ribozymes; a distinct class of metalloenzymes Pyle AM
  16. Biochemistry v.5 An isotopic assay for thymidylate synthase Roberts D
  17. Korean J Microbiol v.31 Functional role of interal guide sequence in splicing activity of T4 thymidylate synthase gene in vivo ShinS;ParkIK
  18. Nucleic Acids Res v.21 Self-cleavage activity of the genomic HDV ribozyme in the presence of various divalent metal ions Suh YM;Kumar PKR;Taira K;Nishikawa S
  19. Dongguk University J Natural Science v.15 Effects of $Mn^{2+}$and $Zn^{2+}$on in vitro RNA splicing of $T_4 Phage thymidylate synthase gene SungJS;ShinS;ParkIK
  20. J Microbiol v.33 Role of $Mg^{2+}$in RNA splicing of $T_4$td intron SungJs;ShinS;ParkIK
  21. J Microbiol v.34 Effects of $K^{+}$ion on in vitro RNA splicing of $T_4$phage thymidylate synthase gene SungJS;ParkIK
  22. Trends in biochem Sci v.14 Self-cleavage of RNA in the replication of small pathogens of plants and animals Symons RH