Preferential activation of the 8-17 deoxyribozyme by Ca2+ ions -: Evidence for the identity of 8-17 with the catalytic domain of the MG5 deoxyribozyme

被引:105
作者
Peracchi, A [1 ]
机构
[1] Univ Parma, Inst Biochem Sci, I-43100 Parma, Italy
关键词
D O I
10.1074/jbc.275.16.11693
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The 8-17 deoxyribozyme is a small RNA-cleaving DNA molecule of potential therapeutic interest. Here, the cleavage rates of 16 variants of the 8-17 deoxyribozyme were measured in the presence of different divalent metal ions. Despite the fact that 8-17 was originally selected in vitro for activity in the presence of Mg2+ (Santoro, S. W., and Joyce, G. F. (1997) Proc. Natl. Acad. Sci. U.S. A. 94, 4262-4266) nearly all the 8-17 variants exhibited substantially higher (up to 20-fold) reaction rates in Ca2+ as compared with Mg2+, This preference for calcium ions critically depended on the nucleoside residues at two specific positions of the deoxyribozyme core. The Ca2+ specificity of 8-17 is strongly reminiscent of the properties of Mg5, an RNA phosphodiester-cleaving deoxyribozyme previously isolated by Faulhammer and Famulok (Faulhammer, D., and Famulok, M. (1996) Angew. Chem. Int. Ed. Engl. 35, 2837-2841). Indeed, analysis of the Mg5 sequence revealed the presence of a complete 8-17 motif, coincident with the conserved region of Mg5, An 8-17 deoxyribozyme modeled after the Mg5 conserved region displayed catalytic features comparable with those reported for the full-length Mg5 deoxyribozyme.
引用
收藏
页码:11693 / 11697
页数:5
相关论文
共 29 条
  • [1] A REEXAMINATION OF MG-25(2+) NMR IN DNA SOLUTION - SITE HETEROGENEITY AND CATION COMPETITION EFFECTS
    BRAUNLIN, WH
    NORDENSKIOLD, L
    DRAKENBERG, T
    [J]. BIOPOLYMERS, 1991, 31 (11) : 1343 - 1346
  • [2] DNA enzymes
    Breaker, RR
    [J]. NATURE BIOTECHNOLOGY, 1997, 15 (05) : 427 - 431
  • [3] A DNA ENZYME WITH MG2+-DEPENDENT RNA PHOSPHOESTERASE ACTIVITY
    BREAKER, RR
    JOYCE, GF
    [J]. CHEMISTRY & BIOLOGY, 1995, 2 (10): : 655 - 660
  • [4] Target site selection for an RNA-cleaving catalytic DNA
    Cairns, MJ
    Hopkins, TM
    Witherington, C
    Wang, L
    Sun, LQ
    [J]. NATURE BIOTECHNOLOGY, 1999, 17 (05) : 480 - 486
  • [5] EVIDENCE FOR THE ROLE OF SOLVATED METAL HYDROXIDE IN THE HAMMERHEAD CLEAVAGE MECHANISM
    DAHM, SC
    DERRICK, WB
    UHLENBECK, OC
    [J]. BIOCHEMISTRY, 1993, 32 (48) : 13040 - 13045
  • [6] ROLE OF DIVALENT METAL-IONS IN THE HAMMERHEAD RNA CLEAVAGE REACTION
    DAHM, SC
    UHLENBECK, OC
    [J]. BIOCHEMISTRY, 1991, 30 (39) : 9464 - 9469
  • [7] Characterization and divalent metal-ion dependence of in vitro selected deoxyribozymes which cleave DNA/RNA chimeric oligonucleotides
    Faulhammer, D
    Famulok, M
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1997, 269 (02) : 188 - 202
  • [8] The Ca2+ ion as a cofactor for a novel RNA-cleaving deoxyribozyme
    Faulhammer, D
    Famulok, M
    [J]. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION IN ENGLISH, 1996, 35 (23-24): : 2837 - 2841
  • [9] Inhibition of the hammerhead ribozyme cleavage reaction by site-specific binding of Tb(III)
    Feig, AL
    Scott, WG
    Uhlenbeck, OC
    [J]. SCIENCE, 1998, 279 (5347) : 81 - 84
  • [10] Frausto da Silva J. J. R., 1991, BIOL CHEM ELEMENTS, P241